Distinct Expression Patterns Of Causative Genes ... - Nature

Distinct Expression Patterns Of Causative Genes Responsible For Hereditary Progressive Hearing Loss In Non-Human Primate Cochlea

Makoto Hosoya1, Masato Fujioka1, Kaoru Ogawa1, and Hideyuki Okano2,*

1 Keio University School of Medicine, Department of Otorhinolaryngology, Head and Neck Surgery, 35

Shinanomachi Shinjyuku-ku Tokyo, 160-8582, Japan

2 Keio University School of Medicine, Department of Physiology, 35 Shinanomachi Shinjyuku-ku

Tokyo, 160-8582, Japan

*[email protected]

Supplementary Figure S.1 The common marmoset cochlea The common marmoset cochlea has 2 and 3/4 turns (between allows) with apical, middle and basal turns as similar to that of human. Note that the cochlear bone capsule was removed.

Supplementary Figure. S2 Cytoplasmic expression of GRHL2 Immunostaining for GRHL2 was observed in the nuclear and cytoplasm. We confirmed the localization by using two different antibodies. Like as shown in Fig.4 (using antibody from Sigma-Aldrich), the immunoreactivity was also detected in cytoplasm with another anti GRHL2 antibody from LSBio. The nuclei were counterstained with Hoechst (blue). Scale bar: 100 µm.

Supplementary Figure. S3 Expressions of CX31, CRYM and GRHL2 in the skin, DFNA5 in the small intestine and ATP6B1 in the kidney For validating antibodies specificities, we performed immunostaining of the skin, the small intestine and the kidney of common marmoset. (a-c) CX31, CRYM and GRHL2 immunoreactivities were detected in hair follicles (HF) or epidermal layers (ED) and not detected in surrounding connective tissues. (d) DFNA5 immunoreactivities were detected in villus (VI) and not detected in crypt (CR) of the small intestine. (e) ATP6B1 immunoreactivities were detected in collecting ducts and tubules and no immunoreactivities was detected in glomeruli (*). The nuclei were counterstained with Hoechst (blue). Scale bar: 100 µm.

Supplementary Figure. S4 Antibody absorption test by antigen peptide For validating antibodies specificities, we performed antibody absorption test by antigen peptide. Immunoreactivities (a and c) were abolished by incubations with antigen peptide (b and d). The nuclei were counterstained with Hoechst (blue). Scale bar: 100 µm.

Supplementary Figure. S5 Validations of secondary antibodies. For validating secondary antibodies’ specificities, we performed immunostaining without primary antibody incubations. No immunoreactivities were observed. The nuclei were counterstained with Hoechst (blue). Scale bar: 100 µm. ms: anti-mouse IgG rb: anti-rabbit Ig G gt: anti-goat IgG, 488: Alexa 488, 555: Alexa 555, 647: Alexa 647

Supplementary Figure. S6 a. CX31 Homo sapiens Callithrix jacchus Mus musculus

MD WK T L QA L L S GV N K Y S T A F GR I WL S V V F V F WV L V Y V V A A E R V WGD E QK D MD WK T L QA L L S GV N K Y S T A F GR I WL S V V F V F R V L V Y V V A A E R V WGD E QK D MD WK K L QD L L S GV N QY S T A F GR I WL S V V F V F R V L V Y V V A A E R V WGD E QK D

50 50 50

Homo sapiens Callithrix jacchus Mus musculus

F D C N T K QP GC T N V C Y D N Y F P I S N I R L WA L QL I F V T C P S L L V I L H V A Y R E E F D C N T K QP GC T N V C Y D N F F P I S N I R L WA L QL I F V T C P S L L V I L H V A Y R E E F D C N T R QP GC T N V C Y D N F F P I S N I R L WA L QL I F V T C P S ML V I L H V A Y R E E

100 100 100


R E R R H R Q K H G D Q C A K L Y D N A G K K H G G L W W T Y L F S L I F K L I I E F L F L Y L L H 150 R E R R H R Q K H G D Q C S K L Y D N A G K K H G G L W W T Y L F S L I F K L I I E F L F L Y L L H 150 R E R K H R Q K H G E Q C A K L Y S H P G K K H G G L W W T Y L F S L I F K L I I E L V F L Y V L H 150


T L W H G F N M P R L V Q C A N V A P C P N I V D C Y I A R P T E K K I F T Y F M V G A S A V C I V 200 T L W H G F D M P R L V Q C A N V A P C P N I V D C Y I A R P T E K K I F T Y F M V G A S A V C I V 200 T L W H G F T M P R L V Q C A S I V P C P N T V D C Y I A R P T E K K V F T Y F M V G A S A V C I I 200


L T I C E L C Y L I C H R V L R G L H K D K P R G G C S P S S S A S R A S T C R C H H K L V E A G E 250 L T I C E L C Y L I C H R V L R A L R K D K P Q G G H T P S S F A S R A S T C R C H H K L V E A G E 250 L T I C E I C Y L I F H R I M R G I S K G K S T K S I S S P K S S S R A S T C R C H H K L L E S G D 250


V D P D P GN N K L QA S A P N L T P I L D P H P GN N K L QA S A P N MT P I P E A D P A S E K L QA S A P S L T P I

270 270 270

b. CRYM Homo sapiens Callithrix jacchus Mus musculus

MS R V P A F L S A A E V E E H L R S S S L L I P P L E T A L A N F S S GP E GGV MQP V R T V V MS R V P A F L S A A E V E E H L R S S S L L I P P L E T A L A N F S S GP E GGV MQP V R T V V MK R A P A F L S A E E V QD H L R S S S L L I P P L E A A L A N F S K GP D GGV MQP V R T V V

50 50 50


P V T K H R GY L GV MP A Y S A A E D A L T T K L V T F Y E D R GI T S V V P S H QA T V L L F E P V T K H R GY L GV MP A Y S A A E D A L T T K L V T F Y E D R GI T S I V P S H QA T V L L F E P V A K H R GF L GV MP A Y S A A E D A L T T K L V T F Y E GH S N T - A V P S H QA S V L L F D

100 100 99


P S N GT L L A V MD GN V I T A K R T A A V S A I A T K F L K P P S S E V L CI L GA GV QA Y S P S N GT L L A V MD GS V I T A K R T A A V S A I A T K F L K P P S S E V L CI L GA GV QA Y S P S N GS L L A V MD GN V I T A K R T A A V S A I A T K L L K P P GS D V L CI L GA GV QA Y S

150 150 149


H Y E I F T E QF S F K E V R I WN R T K E N A E K F A D T V QGE V R V C S S V QE A V A GA D V H Y E I F T E QF S F K E V R I WN R T K E N A E K F A D T V QGE V R V C S S V QE A V T GA D V H Y E I F T E QF S F K E V R MWN R T R E N A E K F A S T V QGD V R V C S S V QE A V T GA D V

200 200 199


I I T V T L A T E P I L F GE WV K P GA H I N A V GA S R P D WR E L D D E L MK E A V L Y V D S I I T V T L A T E P I L F GE WV K P GA H I N A V GA S R P D WR E L D D E L MK E A V L Y V D S I I T V T MA T E P I L F GE WV K P GA H I N A V GA S R P D WR E L D D E L MR QA V L Y V D S

250 250 249


QE A A L K E S GD V L L S GA E I F A E L GE V I K GV K P A H C E K T T V F K S L GMA V E D T QE A A L K E S GD V L L S GA E I F A E L GE V I K GV K P A H C E K T T V F K S L GMA V E D T R E A A L K E S GD V L L S GA D I F A E L GE V I S GA K P A H C E K T T V F K S L GMA V E D L

300 300 299


V A A K L I Y D S WS S GK V A A K L I Y D S WS S GK V A A K L V Y D S WS S GK

314 314 313

c. GRHL2 Homo sapiens Callithrix jacchus Mus musculus

T L E A T K S L RQ K Q G E GPMT Y L N K GQ F Y A I T L S E T GD N K C F R H P I S K V R S V V T L E A T K S L RQ K Q G E GPMT Y L N K GQ F Y A I T L S E T GD N K C F R H P I S K V R S V V T L E A T K S L RQ K Q G E GPMT Y L N K GQ F Y A I T L S E T GD N K C F R H P I S K V R S V V

300 300 300


MV V F S E D K N R D EQ L K Y WK Y WH S RQ H T A K Q R V L D I A D Y K E S F N T I GN I E E I MV V F S E D K N R D EQ L K Y WK Y WH S RQ H T A K Q R V L D I A D Y K E S F N T I GN I E E I MV V F S E D K N R D EQ L K Y WK Y WH S RQ H T A K Q R V L D I A D Y K E S F N T I GN I E E I

350 350 350


A Y N A V S F T WD V N E E A K I F I T V N C L S T D F S SQ K GV K G L P LM I Q I D T Y S Y N N A Y N A V S F T WD V N E E A K I F I T V N C L S T D F S SQ K GV K G L P LM I Q I D T Y S Y N N A Y N A V S F T WD V N E E A K I F I T V N C L S T D F S SQ K GV K G L P LM I Q I D T Y S Y N N

400 400 400


R S N K P I H R A Y CQ I K V F C D K GA E R K I R D E E R K Q N R K K GK GQ A SQ T Q C N S S S R S N K P I H R A Y CQ I K V F C D K GA E R K I R D E E R K Q N R K K GK GQ T SQ T Q C N S S S R S N K P I H R A Y CQ I K V F C D K GA E R K I R D E E R K Q N R K K GK GQ A SQ AQ C N N S S

450 450 450


D GK L A A I P LQ K K S D I T Y F K T MP D L H SQ P V L F I P D V H F A N LQ R T GQ V Y Y N T D GK L A A I P LQ K K S D I T Y F K T MP D L H SQ P V L F I P D V H F A N LQ R T GQ V Y Y N T D GK MA A I P LQ K K S D I T Y F K T MP D L H SQ P V L F I P D V H F A N LQ R T GQ V Y Y N T

500 500 500


D D E R E GGS V L V K RM F R PM E E E F GP V P S K QMK E E GT K R V L L Y V R K E T D D V F D D E R E G - S V L V K RM F R PM E E E F GP V P S K QMK E E GT K R V L L Y V R K E T D D V F D D E R E GS S V L V K RM F R PM E E E F GP T P S K Q I K E E N V K R V L L Y V R K E N D D V F

550 549 550


D A LM L K S P T V K G LM E A I S E K Y G L P V E K I A K L Y K K S K K G I L V NMD D N I I E H D A LM L K S P T V K G LM E A I S E K Y G L P V E K I A K L Y K K S K K G I L V NMD D N I I E H D A LM L K S P T V K G LM E A L S E K Y G L P V E K I T K L Y K K S K K G I L V NMD D N I I E H

600 599 600


Y S N E D T F I L NM E SMV E G F K V T LM E I Y S N E D T F I L NM E SMV E G F K V T LM E I Y S N E D T F I L NM E SMV E G F K I T LM E I

625 624 625

d. DFNA5 Homo sapiens Callithrix jacchus Mus musculus

MF A K A T R N F L R E V D A D GD L I A V S N L N D S D K L QL L S L V T K K K R F WC WQR P K MF A K A T K N F L R E V D A D GD L I A V S N L N D S D K L QL L S L V T K K K R F WC WQR P K MF A K A T R N F L K E V D A GGD L I S V S H L N D S D K L QL L S L V T K K K R Y WC WQR P K

50 50 50


Y QF L S L T L GD V L I E D QF P S P V V V E S D F V K Y E GK F A N H V S GT L E T A L GK V K Y QF L S I T L GD V L K E D QF P S P V V V E S D F V K Y E S K F A N H V S GT L D T A L GK V K Y QI L S A T L E D V L T E GH C L S P V V V E S D F V K Y E S K C E N H K S GA I GT V V GK V K

100 100 100


L N L GGS S R V E S QS S F GT L R K QE V D L QQL I R D S A E R T I N L R N P V L QQV L E G L N V GGS S R V E S QS S F GT L R K QE V D L QQL I R D S A E R T I N L R N P V L QQV L E E L N V GGK GV V E S H S S F GT L R K QE V D V QQL I QD A V K R T V N MD N L V L QQV L E S

150 150 150


R N E V L C V L T Q K I T T MQ K C V I S E HMQ V E E K C GG I V G I Q T K T VQ V S A T E D GN R N E V L C V L T Q K I T T MQ K C V I S E H T Q V E E K C GGMV G I Q T K T VQ V S A T E D GN R N E V L C V L T Q K I MT T Q K C V I S E H VQ S E E T C GGMV G I Q T K T I Q V S A T E D GT

200 200 200


V T K D S N V V L E I P A A T T I A Y GV I E L Y V K L D G L F E F C L L R GK Q GG F E N K K R I VMK D S N V V L E I P A A T T I A Y GV I E L Y V K L D GH F E F C L LQ GK E S G F E N K R R I V T T D T N V V L E I P A A T T I A Y G I M E L F V K Q D GQ F E F C L LQ GK H GG F E H E R K L

250 250 250


D S V Y L D H L V F R E F A F I DMP D A A H G I S SQ D GP L S V L K Q A T L L L E R N F H P F A D S V Y L D P L A F R E F A F I DMP D A GH G I S SQ D GP L S V L K Q A T P L L E R N F H P F V D S V Y L D P L A Y R E F A F L DMP D GGQ G I S SQ D GP L R V V K Q A T L H L E R S F H P F A

300 300 300


E L P E PQQ T A L S D I FQ A V V F D D E L LMV L E P V C D D L V S G L S P T V A V L G - - E L E L P E PQQ T A L S D I FQ A V L F D N E L LMV L E P V C D D V V S G L S P P L V V L G - - E L V L P AQQQ R A L F C V LQ K I L F D E E L L R A L EQ V C D D V A GG LWS SQ A V L AM E E L

348 348 350


K P RQQQ D L V A F LQ L V GC S LQ GGC P GP E D A GS K Q - L FMT A Y F L V S A L A EMP MP RQQQ D VMA F LQ L V GC S LQ G E R P GP E D A GS K R - L F T T A Y F L V S A L A EMP T D SQQQ D L T A F LQ L V GY R I Q G E H P GPQ D E V S NQ K L F A T A Y F L V S A L A EMP

397 397 400


D S A A A L L GT C C K LQ I I P T L C H L L R A L S D D GV S D L E D P T L T P L K D T E R F G I D N A A A L L GT C Y K LQ I I P T L C H L L R A L S D D GV S D L E D P T L A P L K D T E R F G I D N A T V F L GT C C K L H V I S S L C C L L H A L S D D S V C D F H N P T L A P L R D T E R F G I

447 447 450


VQ R L F A S A D I S L E R L K S S V K A V I L K D S K V F P L L L C I T L N G L C A L GR E H S VQ R L F A S A D I T L E R L K S S V K A V I L K D S E I F P L L L C I T L N G L C A L GR E H S VQ R L F A S A D I A L E RMQ F S A K A T I L K D S C I F P L I L H I T L S G L S T L S K E H E E

496 496 500


- - - - - - - - - - - - - - - - - - - - - - E L CQ S GH A T GQ D

512

e. ATP6B1 Homo sapiens Callithrix jacchus Mus musculus

MAM E I D S R P GG L P GS S C N L GA A R E HMQ A V T R N Y I T H P R V T Y R T V C S V N GP MAM E I D S R P GG L P GS G F K L GA A R E HMQ A V T R N Y I T H P R V T Y R T V C S V N GP MA T T V D S R S S G F T GN S C D P GT AQ E H VQ A V T R N Y I T H P R V T Y R T V C S V N GP

50 50 50


L V V L D R V K F AQ Y A E I V H F T L P D GT Q R S GQ V L E V A GT K A I VQ V F E GT S G I D L V V L DQ V K F AQ Y A E I V H F T L P D GT Q R S GQ V L E V A GT K A V VQ V F E GT S G I D L V V L DQ V K F AQ Y A E I V N F T L P D GT Q R S GQ V L E V A GT K A I VQ V F E GT S G I D

100 100 100


A R K T T C E F T GD I L R T P V S E DM L GR V F N GS GK P I D K GP V VMA E D F L D I N GQ A R K T T C E F T GD I L R T P V S E DM L GR V F N GS GK P I D K GP V VMA E D F L D I N GQ SQ K T T C E F T GD I L R T P V S E DM L GR I F N GS GK P I D K GP A VMA E E F L D I N GQ

150 150 150


P I N P H S R I Y P E EM I Q T G I S P I D VMN S I A R GQ K I P I F S A A G L P H N E I A AQ I P I N P H S R I Y P E EM I Q T G I S P I D VMN S I A R GQ K I P I F S A A G L P H N E I A AQ I P I N P H D R I Y P E EM I Q T G I S P I D VMN S I A R GQ K I P I F S A A G L P H N E I A AQ I

200 200 200


C RQ A G L V K K S K A V L D Y H D D N F A I V F A AMGV NM E T A R F F K S D F EQ N GT MGN C RQ A G L V K K S K A V L D Y H D D N F A I V F A AMGV NM E T A R F F K S D F EQ N GT MGN C RQ A G L V K K S K A V L D Y H E D N F A I V F A AMGV NM E T A R F F K S D F EQ N GT MGN

250 250 250


V C L F L N L A N D P T I E R I I T P R L A L T T A E F L A Y Q C E K H V L V I L T DMS S Y A E A V C L F L N L A N D P T I E R I I T P R L A L T T A E F L A Y Q C E K H V L V I L T DMS S Y A E A V C L F L N L A N D P T I E R I I T P R L A L T T A E F L A Y Q C E K H V L V I L T DMS S Y A E A

300 300 300


L R E V S A A R E E V P GR R G F P GY MY T D L A T I Y E R A GR V E GR GGS I T Q I P I L T M L R E V S A A R E E V P GR R G F P GY MY T D L A T I Y E R A GR V E GR GGS I T Q I P I L T M L R E V S A A R E E V P GR R G F P GY MY T D L A T I Y E R A GR V E GR GGS I T Q I P I L T M

350 350 350


P N D D I T H P I P D L T G F I T E GQ I Y V D RQ L H N RQ I Y P P I N V L P S L S R LMK S A I P N D D I T H P I P D L T G F I T E GQ I Y V D RQ L H N RQ I Y P P I N V L P S L S R LMK S A I P N D D I T H P I P D L T G F I T E GQ I Y V D RQ L H N RQ V Y P P I N V L P S L S R LMK S A I

400 400 400


G E GMT R K D H GD V S NQ L Y A C Y A I GK D VQ AMK A V V G E E A L T S E D L L Y L E F LQ G E GMT R K D H GD V S NQ L Y A C Y A I GK D VQ AMK A V V G E E A L T S E D L L Y L E F LQ G E GMT R K D H GD V S NQ L Y A C Y A I GK D VQ AMK A V V G E E A L T S E D L L Y L E F LQ

450 450 450


K F E K N F I NQ GP Y E N R S V F E S L D L GWK L L R I F P K EM L K R I PQ A V I D E F Y S R K F E K N F I NQ GP Y E N R S V F E S L D L GWK L L R I F P K EM L K R I PQ AM I D E F Y S R K F E K N F I T Q GP Y E N R T V F E S L D L GWK L L R I F P K EM L K R I PQ SMT D E F Y S R

500 500 500


E GA LQ D L A P D T A L E GA PQ D P A P D T A L Q GAQQ D P A S D T A L

513 513 513

f. COCH Homo sapiens Callithrix jacchus Mus musculus

MS A AW I P A L G L GV C L L L L P GP - - A GS E GA A P I A I T C F T R G L D I R K E K A D V MS A AW I Q T L F L GV C LQ L L P GP - - MGS E GA A P I A I T C F T R G L D I R K E K A D V MP S S R I P A L C L GAW L L L L L L P R F A R A E GA V P I P V T C F T R G L D I R K E K A D V

48 48 50


L C P GGC P L E E F S V Y GN I V Y A S V S S I C GA A V H R GV I S N S GGP V R V Y S L P GR L C P GGC P L E E F S V Y GN I V Y A S V S S I C GA A V H R GV I S N S GGP V R V Y S L P GR L C P GGC S L E E F S V F GN I V Y A S V S S I C GA A V H R GV I GT S GGP V R V Y S L P GR

98 98 100


E N Y S S V D A N G I Q SQM L S RWS A S F T V T K GK S S T Q E A T GQ A V S T A H P P T GK R E N Y S S V D A N G I Q SQM L S RWS A S F T V T K GK S S T Q E A T GQ A V S T A H P P T GK R E N Y S S V D A N G I Q SQM L S RWS A S F A V T K GK S S T Q E A T GR A V S T A H P P S GK R

148 148 150


L K K T P E K K T G N K D C K A D I A F L I D G S F N I G Q R R F N L Q K N F V G K V A L M L G I G 198 L K K T P E K K T G N K D C K A D I A F L I D G S F N I G Q R R F N L Q K N F V G K V A L M L G I G 198 L K K T P E K K T G N K D C K A D I A F L I D G S F N I G Q R R F N L Q K N F V G K V A L M L G I G 200


T E GP H V G L VQ A S E H P K I E F Y L K N F T S A K D V L F A I K E V G F R GGN S N T GK A L T E GP H V G L VQ A S E H P K I E F Y L K N F T S A K D V L F A I K E V G F R GGN S N T GK A L T E GP H V G L VQ A S E H P K I E F Y L K N F T S A K D V L F A I K E V G F R GGN S N T GK A L

248 248 250


K H T AQ K F F T V D A GV R K G I P K V V V V F I D GWP S D D I E E A G I V A R E F GV N V F I K H T AQ K F F T V D A GV R K G I P K V V V V F I D GWP S D D I E E A G I V A R E F GV N V F I K H T AQ K F F T A D T GV R K G I P K V V V V F I D GWP S D D I E E A G I V A R E F GV N V F I

298 298 300


V S V A K P I P E E L GMVQ D V T F V D K A V C R N N G F F S Y HMP NW F GT T K Y V K P L VQ V S V A K P I P E E L GMVQ D V S F V D K A V C R N N G F F S Y HMP NW F GT T K Y V K P L VQ V S V A K P I P E E L GMVQ D V A F V D K A V C R N N G F F S Y HMP NW F GT T K Y V K P L VQ

348 348 350


K L C T H EQMMC S K T C Y N S V N I A F L I D GS S S V GD S N F R LM L E F V S N I A K T F E K L C T H EQMMC S K T C Y N S V N I A F L I D GS S S V GD S N F R LM L E F V S N I A K T F E K L C T H EQMMC S K T C Y N S V N I A F L I D GS S S V GD S N F R LM L E F V S N I A K T F E

398 398 400


I S D I GA K I A A VQ F T Y DQ R T E F S F T D Y S T K E N V L A V I R N I R Y MS GGT A T GD I S D I GA K I A A VQ F T Y DQ RM E F S F T D Y N T K E N V L A V I R N I R Y MS GGT A T GD I S D I GA K I A A VQ F T Y DQ R T E F S F T D Y N T K E N V L A V L A N I R Y MS GGT A T GD

448 448 450


A I S F T V R N V F GP I R E S P N K N F L V I V T D GQ S Y D D VQ GP A A A A H D A G I T I F S A I S F T V R N V F GP I R E S P N K N F L V I V T D GQ S Y D D VQ GP A A A A H D A G I T I F S A I A F T V R N V F GP I R D S P N K N F L V I V T D GQ S Y D D V R GP A A A A H D A G I T I F S

498 498 500


V GV AWA P L D D L K DMA S K P K E S H A F F T R E F T G L E P I V S D V I R G I C R D F L E S V GV AWA P L D D L K DMA S K P K E S H A F F T R E F T G L E P I V S D V I R G I C R D F L E S V GV AWA P L D D L R DMA S K P K E S H A F F T R E F T G L E P I V S D V I R G I C R D F L E S

548 548 550


QQ QQ QQ

550 550 552

Comparisons of amino-acid sequences between human (Homo sapience), common marmoset (Callithrix jaccus) and mouse (Mus musculus) We compared amino-acid sequences of these three animals and we showed in these six deafness genes amino-acid sequences of common marmoset is more similar to human than mouse. (a) Homology of amino-acid sequences of CX31 was shown. Human vs common marmoset: 95.19%, human vs mouse 82.22% (b) Homology of amino-acid sequences of CRYM was shown. Human vs common marmoset: 99.04%, human vs mouse 88.50% (c) Homology of amino-acid sequences of GRHL2 was shown. Human vs common marmoset: 98.88%, human vs mouse 94.72% (d) Homology of amino-acid sequences of DFNA5 was shown. Human vs common marmoset: 92.14%, human vs mouse 71.37% (e) Homology of amino-acid sequences of ATP6B1 was shown. Human vs common marmoset: 98.44%, human vs mouse 94.72% (f) Homology of amino-acid sequences of COCH was shown. Human vs common marmoset: 98.55%, human vs mouse 93.45%

Supplementary Figure. S7 Immunostaining of mouse specimens. (a) Anti-CX31 antibody recapitulated previous report and strong immunoreactivities were detected in the spiral ligament fibrocytes of mouse and no immunoreactivities was detected in the organ of Corti. (b) Anti-CRYM antibody did not work well against mouse specimens. (c) Anti-GRHL2 antibody used in this report partially recapitulated previous report and immunoreactivities were detected in the organ of Corti. (d) Anti-DFNA5 antibody used in this report partially recapitulated previous report and immunoreactivities were detected in the organ of Corti. (e) Anti-ATP6B1 antibody recapitulated previous report and strong immunoreactivities were detected in the spiral ligament fibrocytes of mouse. (f) Anti-COCH antibody recapitulated previous report and strong immunoreactivities were detected in the spiral ligament fibrocytes of mouse and no immunoreactivities was detected in the organ of Corti. SLF: Spiral Ligament Fibrocytes, *. Organ of Corti. The nuclei were counterstained with Hoechst (blue). Scale bar: 100 µm.

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