Fig 2: Dynamic expression of Kindlin-1 in skin wound healing in mice. Assessment of Kindlin-1 in skin wound healing in mice with or without EGF treatment. (A1)Construction of wound healing model in mice. (A2) Healing rate of EGF(−) and EGF(+) group at the 14th day. *P<0.05. (B1) Kindlin-1 expression was detected by real-time PCR. (C1) Kindlin-1 expression was detected by western blot analysis and with GAPDH as an internal control. (C2) Kindlin-1 expression was quantified and presented by the fold changes compared with the relative control [value of EGF(−) 0 days or EGF(+) 0 days]. *P<0.05, significant difference compared to the normal group [EGF(−) 0 days, EGF(+) 0 days]. #P<0.05.

Fig 3: Distribution of Kindlin-1 in mouse skin tissues. Expression pattern of Kindlin-1 in mouse tissue slices was observed at the indicated time intervals after operation. (A) Normal; (B–D) EGF(−) 3 days, 7 days and 14 days, respectively; (E–G) EGF(+), 3 days, 7 days and 14 days, respectively. MT, migratory tongue; HE, hyperproliferative epithelium. The dotted line indicates the wound edge. Scale bar, 100 µm.

Fig 4: EGF-induced F-actin re-arrangement is disrupted by kindlin-1 shRNA. Staining of F-actin in HaCaT cells was by visualized with phalloidin staining (red) and confocal microscopy, DAPI counterstaining for cell nuclei (blue). (A) Control shRNA-transected HaCaT cells without EGF stimuli. (B) Control shRNA-transected HaCaT cells cultured with EGF for 6 h. (C) Kindlin-1 shRNA-transfected HaCaT cells without EGF stimuli. (D) Kindlin-1 shRNA-transfected HaCaT cells cultured with EGF for 6 h. Scale bar, 10 µm.

Fig 5: Expression of Kindlin-1 in EGF-stimulated HaCaT cells. (A1) Expression of Kindlin-1 in HaCaT cells in response to EGF (10 ng/ml) was assessed by western blot analysis. (A2) Blots were quantified and compared with GAPDH, the graphs show quantification data. Values were reported as the means ± SEM (n=4). *P<0.05, significant difference compared to the value at 0 h. (B–C) Knockdown efficiency of Kindlin-1 protein in HaCaT cells was determined by fluorescence microscopy and western blot analysis. Quantification data are shown in the graph (C). *P<0.05, significant difference compared with the normal group; ΔP<0.05, significant difference compared with the control shRNA group. EGF, epidermal growth factor; N, normal HaCaT cells; C, control shRNA-transfected HaCaT cells; K, Kindlin-1 shRNA-transfected HaCaT cells.