Fig. 1
The levels of CSDE1 protein decrease during hESC differentiation. a Quantitative proteomic analysis of CSDE1 levels comparing H9 hESCs with their NPC and neuronal counterparts. Graph represents the mean (±confidence interval) of relative abundance differences calculated from the log2 of label-free quantification (LFQ) values (hESCs (n = 9), NPCs (n = 6) and neurons (n = 5)). Statistical comparisons were made by limma’s moderated t-test (P-value: **** (P < 0.0001)). b Western blot analysis with antibody to CSDE1. The graph represents the CSDE1 relative percentage values (corrected for β-actin loading control) to H9 hESCs (mean ± s.e.m. of five independent experiments). c Western blot of CSDE1 in H1 hESCs and their differentiated mesoderm and cardiomyocyte counterparts. The graph represents the CSDE1 relative percentage values (corrected for β-actin loading control) to H1 hESCs (mean ± s.e.m. of two independent experiments). d Western blot analysis with antibody to CSDE1. The graph represents the CSDE1 relative percentage values (corrected for β-actin loading control) to H9 hESCs (mean ± s.e.m. of three independent experiments). e Immunocytochemistry of H9 hESCs and NPCs (2 weeks under neural induction treatment) with antibody to CSDE1. Hoechst staining was used as a marker of nuclei. Scale bar represents 20 μm. f Immunocytochemistry of cell cultures at early stages of the neural induction treatment (2 days) with antibody to CSDE1. OCT4 and Hoechst staining were used as markers of pluripotency and nuclei, respectively. These cultures contain undifferentiated (OCT4-positive) and differentiated (OCT4-negative) cells. White arrow indicates OCT4-negative cells. Scale bar represents 20 μm. g Quantitative PCR (qPCR) analysis of CSDE1 mRNA levels. Graph (CSDE1 relative expression to H9 hESCs) represents the mean ± s.e.m. of three independent experiments. h CSDE1 relative expression to H1 hESCs represents the mean ± s.e.m. of three independent experiments with three biological replicates. i CSDE1 relative expression to H9 hESCs represents the mean ± s.e.m. of two independent experiments with three biological replicates. In b–d and g–i, statistical comparisons were made by Student’s t-test for unpaired samples. P-value: *(P < 0.05), **(P < 0.01), **** (P < 0.0001)
