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. 2023 Jun 26;10(24):2300152. doi: 10.1002/advs.202300152

Figure 6.

Figure 6

Optimal level of focal adhesion kinase inhibition improves iN cell generation. A) Reprogramming efficiency of BAM‐transduced fibroblasts cultured in the absence and presence of various concentrations of the FAK inhibitor, PF573228 (denoted as PF in this figure) at day 14 (n = 3). Significance was determined by a one‐way ANOVA and Dunnett's multiple comparison test. B) Immunofluorescent images of TUBB3+ iN cells derived in the presence of 1 µm PF573228 (PF) displaying a typical neuronal morphology and co‐expressing mature neuronal markers, NeuN, MAP2 and synapsin at day 21. Scale bar, 50 µm. C) Representative traces of spontaneous changes in membrane potential in response to current injection from iN cells obtained in the absence and presence of 1 µm PF573228. The inhibitor was administered during the first 7 days of reprogramming. D) Western blot analysis of FAK and mesenchymal marker expression in BAM‐transduced fibroblasts that were treated with various concentrations of PF573228 for 2 days. E) qRT‐PCR analysis of neuronal gene expression at day 5 from non‐transduced (NT) and BAM‐transduced fibroblasts cultured in the absence and presence of 5 µm PF573228 for 4 days (n = 3). Expression level normalized to BAM‐transduced fibroblasts treated with DMSO. Significance determined by two‐tailed, unpaired t‐test, compared to the DMSO condition for transduced cells for the same gene. Bar graphs show mean ± standard deviation (*p < 0.05, **p < 0.01).