Figure 4.

Generation of skin equivalents from CD271 silenced ETA cells. (A) Protein extracts from freshly isolated KSC, ETA, and LTA cells were immunoblotted for CD271 and the relative expression was densitometrically evaluated by Fiji-ImageJ software. (B) Confirmation of CD271 downregulation in protein extracts from scramble or CD271 ETA cells by immunoblotting. (C) Skin equivalents derived from KSC, CD271 silenced ETA, and scrambled ETA cells were paraffin-embedded at 6 and 12 days from cell seeding and sections were stained with H&E. Bar = 100 μm. Epidermal thickness was measured by Fiji-ImageJ software. (D) Sections of skin equivalents obtained from KSC, CD271 silenced ETA, and scrambled ETA cells were stained with anti-Ki67 antibodies by IHC, anti-Survivin, anti-K15, and anti-Involucrin antibodies by IF. Bar = 100 μm. The number of positive cells or stained areas was evaluated by image pixel count using Fiji-ImageJ. Data are represented as mean ± SEM. Ordinary one-way ANOVA followed by Tukey’s multiple comparisons test were performed, and comparisons are indicated. ns: P > .05; * .01 < P < .05; ** P < .01; *** P < .001; ****P < .0001.