Fig 2: Overexpression of PC4 inhibits cell migration and proliferation in vitro. The migration and proliferation of primary fibroblasts derived from PC4+/+ mice decreased. (a) Images of primary fibroblasts after 24 hours from the initial migration test. Yellow bar: 200 μm. (b) Scatter diagram quantifying the relative migration area. Each sample was measured 6 times at the indicated time point. Each experiment was repeated thrice. P values were calculated with t-text. (c) Proliferation was measured by CCK-8 assays. P values were calculated with two-way ANOVA. (d) Clone formation assay was performed with WT and PC4+/+ primary fibroblasts. *p < 0.05, *p < 0.01. PC4 human positive cofactor 4, PC4+/+ PC4 knock-in mouse model, CCK-8 cell counting kit-8, ANOVA analysis of variance

Fig 3: Overexpression of PC4 reduces angiogenesis and cell migration in wounds. (a) CD31 (red) and DAPI (blue) expression was shown in the representative images of WT and PC4+/+ mice after 0–14 days after trauma. Scale bar = 100 μm. Arrows show representative CD31-positive/DAPI-positive cells. (b) α-SMA (red) and DAPI (blue) expression was shown in the representative images of WT and PC4+/+ mice after 0 to 14 days after trauma. Scale bar = 100 μm. Arrows show representative α-SMA-positive/DAPI-positive cells. PC4 human positive cofactor 4, WT wild-type, α-SMA alpha smooth muscle actin

Fig 4: Overexpression of PC4 inhibits wound healing in vivo. (a) Schematic of the wound healing model. (b) Representative images of the skin wound healing for the indicated times (n = 8 per group). (c) Quantification of the wound areas was performed using Image J software. Data represented the means ± SD, *p < 0.05, **p < 0.01. P values were calculated with two-way ANOVA. (d) Pathological studies confirmed that delayed healing in PC4+/+ mice during the granulation tissue maturation process. Representative images of hematoxylin and eosin-stained skin sections from biopsies on days 3, 7, 10 and 14 in WT and PC4+/+ mice; n = 3 mice per group. Scale bar = 200 μm. PC4 human positive cofactor 4, SD standard deviation, ANOVA analysis of variance, PC4+/+ PC4 knock-in mouse model, WT wild-type

Fig 5: Overexpression of PC4 inhibits cell proliferation in wounds (a) Ki67 immunofluorescence staining revealed cell proliferation at the healing sites of skin wounds on days 0, 3, 7, 10 and 14. Scale bar = 100 μm. Arrows show representative Ki67-positive/DAPI-positive cells. (b) The relative quantitative changes in Ki67-positive cells were observed in the skin wound healing sites after 0, 3, 7, 10 and 14 days in WT and PC4+/+ mice. *p < 0.05, **p < 0.01. P values were calculated with two-way ANOVA. PC4 human positive cofactor 4, WT wild-type, ANOVA analysis of variance