Comparison of CYP gene number in Insecta, Crustacea and T. urticae. CYP2 clan mitochondrial CYP clan. CYP3 clan. CYP4 clan. Total. Insecta. Drosophila.
mouse pri-miR-141. 185 bp mouse pri-miR-200a. 201 mouse pri-miR-429. 170 bp mouse pri-miR-2000. 284 bp. Mouse Zeb2. 223 bp mouse pri-miR-200b.
$2,400. $99,776. Oligo Pool amplification with. Kapa HiFi Uracil+. $24. -. USER treatment + End Repair. $36. -. Assembly PCR. $12. -. Sequence Verification.
Parental B16 and B16mCAR (with forced expression of murine CAR) were ... activity. (B) B16 cells were transduced with AdRGD-PG-eGFP (MOI=50), treated or.
Figure S5. Genotyping analysis of C2C12 cells transfected with either donor. (mCherry-puromycinR) only, TALENs only, or donor plus TALENs. (A) A schematic.
MMP9. Matrix metallopeptidase 9 (gelatinase B, 92kDa gelatinase, 92kDa type IV collagenase). E12 NM_014071. NCOA6. Nuclear receptor coactivator 6. F1.
B. Digital panning of a diverse donor-17 scFv library against a clade-C V1V2-ferritin nanoparticle (negative control). H. 28,145. 26,448. 16.5%. Pan0. 171,308.
Different letters denote significant differences according to 3-way ANOVA and Tukey's test. (p
Table S1. Primers used for PCR and construct generating. Primers for real-time PCR Sense primer (5’-3’) pri-miR-125b-1 pri-miR-125b-2 pri-miR-21 pri-miR-23b-27b-24-1 pri-miR-30b
Primers for Chromatin Immunoprecipitation mir-17-92 (-1733 to -1583) mir-17-92 (-1484 to -1297) mir-17-92 (-947 to -772) mir-130a (-3095 to -2912) mir-23b-27b-24-1 (-1396 to -1209)
mir-125b-1 (-2523 to -2334) mir-125b-1 (-1113 to -923) mir-21 (+1124 to +1305) mir-21 (+1340 to +1534) mir-30b (-415 to -231) IL-8 (-92 to +90)
CCTTAAAATGCAACCTACTAA
ACAAGCAACTATTTTCTGTGA
TGAGATGAATAACTCCTGCT
ATTTTACGTTACTTTTGGTCG
CACGAACCAGTGATATGTGCT
AAGAAGCCCAATCAGGAGC
TTCTGACTGTCCTTGGGCA AGCAGCTAGCAGGGTGATGT
AAGGGAGCTGAGATACACCCA CATGGGAAGAACAGAGGATGA
CGTCCATAAAGAAAGGCCAC
CGCAAGACCTAGAGATCAGG
TCATCTTCCCATCTGCCT
CTGCGGATTCTTTGAAGC
GGAGTGGATGGGTTCTGCCTTA
CAAGGTGGATTGCATCGAGG
TGCAACAGACTGGCCTTC
CATGCAAGACTGTTATCCAATCT
GGGAAGGATATAGGAAGGCTGG GGGCCATCAGTTGCAAATC
Primers for Promoter constructsa mir-17-92 tacgcgtAGGAAGTATGAGCGAAACCC (-1747 to -790 )
GCCCAGGCTAGTCACAAACA GGAAGAAACCACCGGAAGGAA
taagcttAGCACCTCCTTCTTCACATTG
mir-17-92
tacgcgtGTTGCTTGTATGTCAGATTTCC
taagcttAGCACCTCCTTCTTCACATTG
mir-17-92
tacgctGACATATTAGTCAATGCCGACC
taagcttAGCACCTCCTTCTTCACATTG
mir-17-92
(-1747 to -1328 )
tacgcgtAGGAAGTATGAGCGAAACCC
taagcttGGTCGGCATTGACTAATATGTC
mir-17-92
tacgcgtAGGAAGTATGAGCGAAACCC
taagcttGGAAATCTGACATACAAGCAAC
(-1583 to -790) (-1328 to -790)
(-1747 to -1583)
mir-17-92
taagcttGGTCGGCATTGACTAATATGTC
(-1583 to -1328)
tacgcgtGTTGCTTGTATGTCAGATTTCC
mir-130a mir-125b-1
tacgcgtTCATGAGCGAACACATCAACC
tgctagcTTCGGGAAGCTCTCGATACCT
tacgcgtAGAAAGGCCACCAAGATTCAC
tgctagcTGAGAGGAGCGCAACAATGT
mir-125b-1
(-1129 to +106)
tacgcgtAAAGGGTCATCTTCCCATCTG
tgctagcTGAGAGGAGCGCAACAATGT
mir-23b-27b-24-1
tacgctGCAGCTAGCAGGGTGATGTT
(-2514 to +106)
(-1396 to +41)
mir-30b mir-21
(-332 to +1957)
mir-21
(+1269 to +1957)
tctcgagACAGGGAGCGAACAGGTTA
tacgcgtTTAATTCTGGATGCCCTTGCT
tgctagcTTGCCCAGGCTAGTCACAA
tacgcgtGAAGTTGGTTTGCCAGTGTTCC
tgctagcTCCCAGAGGTGCCATTTAGC
tacgcgtATCCACCCTCGATGCAATC
tgctagcTCCCAGAGGTGCCATTTAGC
Listed in this table are all the primers used in this study for the real-time PCR and ChIP analyses and construct generating. aRestriction enzyme sites were indicated by lowercase letters.
Table S2. miRNA expression profile in biliary epithelial cells following LPS stimulation.
Data represent the average of log2 (Hy5/Hy3) ratios from non-stimulated cell cultures (n = 3), LPS treated cultures (n = 3) by using the miRCURYTM LNA Array (Version 8.1).
Figure S1. Promoter binding of p65 transactivates the IL-8 gene in biliary epithelial cells in response to LPS stimulation. (A) p65-dependent upregulation of IL-8 mRNA in cholangiocytes after LPS treatment. Bars represent the levels of IL-8 mRNA in cells following LPS stimulation in the presence or absence of SC-514 as assessed by real-time PCR. (B) A schematic diagram shows the structure of IL-8 gene. ChIP analysis demonstrated increased binding of p65 to the binding site at IL-8 promoter in cells following stimulation. , p < 0.05 t test vs non-stimulated cells; #, p < 0.05 t test vs LPS-stimulated cells.
Figure S2. Promoter binding of p65 transactivates mir-130a gene in biliary epithelial cells in response to LPS stimulation. (A) The schematic diagram shows one potential binding sites in the putative promoter element of mir-130a gene. ChIP analysis revealed increased binding of
p65 to the promoter binding site in H69 cells following LPS treated. (B) H69 cells were transfected with luciferase reporter construct covering the potential binding sites of the mir-130a promoter and then exposed to LPS in the presence or absence of SC-514. (C) H69 cells were co-transfected with the pCMV-p65 to overexpress p65 and the luciferase reporter construct containing the mir-130a promoter. , p < 0.05 t test vs non-stimulated cells (in B) or empty pCMV vector control (in C); #, p < 0.05 t test vs LPS-stimulated cells (in B).
Figure S3. Promoter binding of p65 transactivates miR-125b-1 gene in biliary epithelial cells in response to LPS stimulation. (A) The schematic diagram shows two potential binding sites in the putative promoter element of mir-125b-1 gene. ChIP analysis revealed increased binding of p65 to the binding site at -1059, but not at -2455, of mir-125b-1 promoter element in cells following stimulation. (B) H69 cells were transfected with various luciferase reporter constructs spanning the potential p65 binding sites of the mir-125b-1 promoter. The transfected cells were exposed to LPS in the presence or absence of SC-514. , p < 0.05 t test vs nonstimulated cells; #, p < 0.05 t test vs LPS-stimulated cells.
Figure S4. Promoter binding of p65 transactivates mir-21 gene in biliary epithelial cells in response to LPS stimulation. (A) The schematic diagram shows two potential binding sites in the putative promoter element of mir-21 gene. ChIP analysis revealed increased binding of p65 to the binding site at +1395 of mir-21 promoter element in cells following stimulation. Although the two potential binding sites for NF-B subunit p65 are adjacent to each other, a stronger signal for the binding site at +1395 was detected. (B) H69 cells were transfected with various luciferase reporter constructs spanning the potential p65 binding sites of mir-21 promoter. The transfected cells were exposed to LPS in the presence or absence of SC-514. , p < 0.05 t test vs non-stimulated cells; #, p < 0.05 t test vs LPS-stimulated cells.
Figure S5.
Promoter binding of p65 transactivates C3orf9 (mir-23b-27b-24-1) gene in
biliary epithelial cells in response to LPS stimulation. (A) The schematic diagram shows one potential binding sites in the putative promoter element of C3orf9 gene. ChIP analysis revealed increased binding of p65 to the binding site at -1254 of C3orf9 promoter element in cells following stimulation. (B) H69 cells were transfected with various luciferase reporter constructs spanning the potential p65 binding sites of the C3orf9 promoter. The transfected cells were exposed to LPS in the presence or absence of SC-514. , p < 0.05 t test vs non-stimulated cells; #, p < 0.05 t test vs LPS-stimulated cells.
Figure S6. Promoter binding of p65 transactivates the mir-30b gene in biliary epithelial cells in response to LPS stimulation. (A) The schematic diagram shows one potential binding sites in the putative promoter element of mir-30b gene. ChIP analysis revealed increased binding of p65 to the binding site at -338 of mir-30b promoter element in cells following stimulation. (B) H69 cells were transfected with various luciferase reporter constructs spanning the potential p65 binding sites of the mir-30b promoter. The transfected cells were exposed to LPS in the presence or absence of SC-514. , p < 0.05 t test vs non-stimulated cells; #, p < 0.05 t test vs LPS-stimulated cells.
FIGURE S1.
IL8 mRNA (relative expression)
A 16 *
12 8
*
4
# #
0
LPS SC-514
-
+
4h -
4h +
8h -
B
8h +
IL8 Chr4 -72
+1
100 bp GTGGAATTTC
Genomic sequence
3.0
%input
Input IP: anti- P65 IP: IgG LPS
2.0 1.0 0
-
+
LPS
-
+
FIGURE S2.
A
mir-130a 1kb Chr11 -2968
+2904
+1
Genomic sequence GGGAATTTGC
%input
3.0
Input IP: anti- P65
2.0 1.0
IP: IgG LPS
0
-
+
LPS
B
+
Control Control+SC-514 LPS stimulation LPS +SC-514
