Fig. 7
Exogenous PELI2 overexpression rescues the effect of ALKBH5 knockdown on re-epithelialization and wound healing in vivo. a Schematic ideogram showing the intradermal injection of Lv-PELI2 and Lv-NC (1 × 107 TU/ml, 1% Lipo3000) into the skin at the wound edge of WT and Alkbh5‒/‒ mice at d0 and d1 after wound establishment. All the mice were sacrificed at PWD8. b The restoration of PELI2 mRNA expression in the skin of the wound edge of Alkbh5‒/‒ mice was confirmed by qRT‒PCR. Five wounds from five mice were included in the analysis. The relative gene expression levels are shown as the mean ± SD. One-way ANOVA, *P < 0.05, **P < 0.01. c IF confirmed the restoration of PELI2 expression in the epidermis of the wound edge of Alkbh5‒/‒ mice after Lv-PELI2 transfection. Dotted lines denote epidermal boundaries. Epi epidermis. Scale bar: 25 μm. d, e Representative images of cutaneous wounds (d) of WT mice transfected with Lv-NC and Alkbh5‒/‒ mice transfected with Lv-PELI2 or Lv-NC on PWD0, PWD2, PWD4, PWD6, and PWD8. Scale bar: 2 mm. The black dashed circle delineates the original wound of 5 mm width. The rate of wound closure (e) was quantified by using ImageJ software and is expressed as the percentage of the nonhealing wound area. Ten wounds of five mice were included in the analysis. The relative percentages of wound closure are shown as the mean ± SD. One-way ANOVA (at 2 days, 4 days, 6 days), ***P < 0.001. Mann–Whitney test (at 8 days), two-sided *P < 0.05 (KO + Lv-PELI2 vs. KO + Lv-NC). f, g Histological analysis (f) of wound re-epithelialization in WT mice transfected with Lv-NC and Alkbh5‒/‒ mice transfected with Lv-PELI2 or Lv-NC at eight days after wounding. The line with arrowheads indicates the 5 mm width of the original wound gap. The line with blunt ends delineates the epithelial gap, indicating the width of the nonepithelialized wound area. Quantification of the percentage of re-epithelialization (g). Ten wounds of five mice were included in the analysis. The relative percentages of re-epithelialized wounds are shown as the mean ± SD. Mann–Whitney test, two-sided **P < 0.01, ****P < 0.0001. Dotted lines denote epidermal boundaries. Scale bar: 200 μm