Figure 5.

The analysis of proliferation and migration capabilities of 4-week-old NT2/A transduced with SOX2 and SOX9 expression vectors. (A) Relative cell viability of 4-week-old NT2/A after transduction with SOX2 or SOX9 expression vectors (indicated as SOX2 OE and SOX9 OE, where OE stands for overexpression) was measured by MTS assay and compared with the viability of cells transduced with the corresponding empty vector (control). Data presented are means ± SEM (n = 3); p-values were calculated using Student’s t-test: * p ≤ 0.05. Representative images of Ki67 immunostaining (green) of 4-week-old NT2/A transduced with SOX2 expression vector or empty vector (control) counterstained with nuclear marker DAPI (blue). The boxed region is magnified to show the nuclear localization of Ki67. Micrographs were taken using a 60× objective. Scale bar—20 µm. The percentage of Ki67+ cells presented in the chart was calculated relative to the total number of DAPI-labeled nuclei in three independent experiments. Data represent means ± SEM, and p-values were calculated using Student’s t-test: ** p ≤ 0.01. (B) Representative phase contrast images of wound closure experiments. The migratory capability of 1-week-old NT2/A and 4-week-old NT2/A transduced with SOX9 expression vector or empty vector (control) was measured 24 h after scratch injury. Scale bar—100 µm. The statistical analysis of the percentage of wound closure was obtained from three independent experiments. Data represent means ± SEM, and p-values were calculated using Student’s t-test: ** p ≤ 0.01, *** p ≤ 0.001.