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mentary Fi. 41 inductio .... Rb α-Caspase-3 (total CASP3). Cell Signaling ... Rb α-GFP. Cell Signaling. 2956. WB (1:1,000). Rb α-GFP. Abcam ab290. IP (1 µg).
SUPPLEMENTARY INFORMATION

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Supplementary Figures

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Proteins with enhanced ubiquitination after LIN41 induction in both LIF and RA conditions 2410127L17Rik Akap2 Atp13a3 Atp5g2 Birc6 Cacybp Capzb Cdk1 Cep55 Cnot1 Csda Cul7 Cyp26a1 Dnd1 Eef1a1 Eif1 Eps15l1 Fam115a Flii

Fzd2 Gtf2h4 H2afx Hist2h2aa2 Hist2h2ac Hspa1a Hspa1b Hspa1l Hspa8 Huwe1 Nisch Nop56 Nsfl1c Otud5 Piwil2 Psmd14 Rabac1 Rnf10 Rpl19

Rpl6 Rps24 Set Slc4a7 Snrpb Snrpn Solh Sumo2 Sumo3 Tapbp Tex11 Trim35 Trim71 Trp53inp1 Trpc4ap Ube3a Ube4b Usp5

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Supplementary Figure 1, related to Figure 1



Table lists those proteins with enhanced ubiquitination in response to LIN41 induction (with



≥2.5 fold) in common between data sets from pluripotent (Dox+ + LIF vs Dox- + LIF) and

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differentiated (Dox+ + RA vs Dox- + RA) conditions in iLin41 cells.

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Supplementary Figure 2, related to Figure 3

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(a) Flag-LIN41 was co-expressed together with negative control GFP, positive control AGO2-

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GFP or p53-GFP in HeLa cells. Western blots of input extracts and bound proteins after

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Flag-IP were probed with indicated antibodies. AGO2-GFP and p53-GFP, but not GFP alone,

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co-purified with LIN41.

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(b) LIN41-GFP was co-transfected together with empty Flag vector, Flag-AGO2 as positive

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control or Flag-p53 in HeLa cells. Input and bound proteins were probed by Western blotting

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with α-GFP or α-Flag antibodies. LIN41 co-immunoprecipitates p53 and AGO2.

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Supplementary Fiigure 3, related to Fig gure 4

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(a) LIN4 41 induction does nott affect prolliferation in cycling ce ells, but incrreases proliferation

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after thrree days of RA-induced d differentia ation. iLin41 1 cells were e cultured unnder prolife erative or

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differentiation conditions. Brd dU incorpo ration was monitored at 24 h iintervals. Data D are

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presentted as mean n values ± SD S of three independent experime ents. * p < 00.05

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(b) LIN N41 induction during neural d ifferentiatio on facilitate es G1-S/G22 transition. Flow

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cytomettric cell cycle analysis was perform med on iLin n41 cells tre eated with R RA for three e days in

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the absence or pre esence of Dox. D A repre esentative histogram h gated for ce lls in G1, S and G2

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is show wn.

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(c) Statistical analyysis of cell cycle c phase e distributio on. Data are e presentedd as means ± SD of

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three independent experimentts. * p