Figure 5.

Evaluation of stemness markers expression in untreated (CTR) and 10 µM CoCl₂-treated cells for 6 h, kept in culture for 24 h or 48 h. (A) Representative quantitative RT-PCR analysis of OCT-4, SOX-2, and NANOG stemness genes. Relative quantification of each target’s gene expression was calculated using the ΔΔCt method and presented as fold change to the CTR 24 h (calibrator). (B) Representative IHC images of SOX-2 and NANOG proteins nuclear localization (green fluorescence, arrows) in CTR and CoCl₂-treated oAECs. Nuclei were counterstained with DAPI (blue fluorescence). Arrowheads indicate negative stemness nuclear localization in the cells. Scale bar: 10 µm. Histograms show the percentage of cells with positive nuclei to SOX-2 or NANOG/100 total cells. Values statistically different for ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. CTR 24 h and ^## p < 0.01, ^#### p < 0.0001 vs. 10 µM 24 h.