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. 2021 Mar 23;22(6):3256. doi: 10.3390/ijms22063256

Figure 1.

Figure 1

Treatment of iPSCs with 0.05 µM BIBR1532 allows continued culture, maintains pluripotency, and does not induce apoptosis; however, 0.05 µM BIBR1532 has no effect on TERT expression throughout iPSC-MN directed differentiation. (A1) Qualitative immunocytochemistry for DAPI, OCT4, and activated CASP3 in iPSCs treated with 0.05 µM BIBR1532 or DMSO for 15 days. Scale bars = 20 µm. (A2) Quantitative analysis of the percentage of OCT4+ nuclei and OCT4+CASP3+ nuclei in 0.05 µM BIBR1532 treated iPSCs and DMSO controls. Data presented as mean ± S.E.M. Unpaired t-test. Three biological repeats (represented by datapoints), five fields per cover slip. (B) qRT-PCR analysis of OCT4 expression at day 0 and day 7 of neural induction, in cells treated with 0.05 µM BIBR1532 or DMSO, normalised over GAPDH expression, relative to day 0 control iPSCs. (C) qRT-PCR analysis of expression of TERT throughout iPSC-MN differentiation in cells treated with 0.05 µM BIBR1532 or DMSO, normalised to GAPDH expression, relative to day 0 control iPSCs. (B,C) Data presented as mean ± S.E.M. One experimental block, three biological replicates. Datapoints represent biological replicates. From day 7 onwards, an additional technical replicate of the Ctrl 1 line was included in controls, represented as an additional datapoint above. Two-way ANOVA, with Tukey’s test for multiple comparisons. n.s. = non-significant, with significance placed at a p-value of <0.05. p-values displayed graphically refer to the statistical difference between the indicated timepoint and day 0 control samples.