Journal: Stem cell research & therapy

Article Title: OCT4 translationally promotes AKT signaling as an RNA-binding protein in stressed pluripotent stem cells.

doi: 10.1186/s13287-025-04229-1

Figure Lengend Snippet: Fig. 1 Post-transcriptional regulation of AKT1 by OCT4. A NCCIT, H1 and H9 cells were transfected with scramble siRNA or OCT4 siRNAs for 48 h, and the mRNA levels of OCT4, NANOG, SOX2 and AKT1 were determined by qRT-PCR. B NCCIT, H1 and H9 cells were transfected with scramble siRNA or OCT4 siRNAs. After 48 h, cells were lysed and the whole cell lysates were subjected to immunoblotting with indicated antibodies. Data shown were from one of two independent experiments that gave similar results, and the densitometric analyses were presented in Supplementary file 6: Fig. S1D and E. C, D H9 cells were treated with 5 ng/ml Actinomycin D (ACTD) for varying period, samples were collected for qRT-PCR (C) and immunoblotting (D). The band intensities of AKT and OCT4 in D were normalized by those of GAPDH, and plotted in (C). Data in (A) and C were presented as mean ± SD of three independent experiments. In C, values at 24 h and 48 h were statistically compared with those at 0 h. Unless otherwise specified, the statistical significance of compared measurements was evaluated with one-way ANOVA and LSD test using SPSS 19.0. ns, not significant, **P < 0.01, ***P < 0.001, ****P < 0.0001. E Relative quantity of OCT4 proteins in cytoplasmic extract (CE), nuclear extraction (NE) and nuclear pellet extraction (NP, enriched in nuclear matrix proteins) of H1 cells. Full-length blots/gels are presented in Supplementary file 7: Fig. S28

Article Snippet: The rabbit monoclonal anti-OCT4 antibody (2890S), anti-pan Akt antibody (4691), anti-Lamin B1 antibody (17416), anti-TBP antibody (44059), anti-Histone H3 antibody (4499), anti-HSP90 (4877) antibody, antiHNRNPA1 antibody (8443) and anti-Tubulin antibody (2148) used in this study were purchased from Cell Signaling Technology.

Techniques: Transfection, Quantitative RT-PCR, Western Blot, Extraction