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. 2020 Feb 5;6(6):eaay0264. doi: 10.1126/sciadv.aay0264

Fig. 5. LINC00458 interacts with SMAD3, a core component of the endoderm-specific transcription factor.

Fig. 5

(A) Immunoblot of SMAD2/3 (SMAD2 (Ser465/467)/SMAD3 (Ser423/425) phosphorylation in hPSCs cultured on soft and/or hard substrates and TCPS. (B) qRT-PCR detection of LINC01356 and LINC00458 pulled down by SMAD2/3- or SMAD1-specific antibodies, as compared to that with immunoglobulin G (IgG), based on the RNA-binding protein immunoprecipitation (RIP) assay. SMAD2/3_S and SMAD2/3_H, soft and hard group samples, respectively, with anti-SMAD2/3 antibody; SMAD1_S and SMAD1_H, soft and hard group samples, respectively, with anti-SMAD1 antibody. (C) Immunoblot detection of the levels of SMAD2/3 in hPSCs on soft substrates transfected with SMAD2 or SMAD3 siRNAs. (D) qRT-PCR detection of LINC00458 and MALAT1 in RIP complex in hPSCs with SMAD2 or SMAD3 siRNAs. (E) A working model of soft substrate induces LINC00458 to interact with SMAD2/3, which promotes endoderm specification. The results are presented as means ± SD of triplicates. **P < 0.005, *P < 0.05, one-way ANOVA (n = 3 independent experiments). NS, not significant.