Fig. 8

Comparison of PRP pre-treatment with AG490, a putative JAK/STAT inhibitor, in deactivating JAK/STAT signalling and alleviating apoptosis in I/R-injured skin flaps. A Experimental protocol: twenty-four mice were randomly and evenly divided into six groups as described in “Materials and methods” section. PBS (green arrows), PRP (yellow arrows), or AG490 (red arrows) was subcutaneously injected to the flaps prior to I/R insult or without I/R insult. Skin flap samples were collected and processed at 16 h on day 0 for Western blotting. Blue rectangles represent the period for model establishment. Gray rectangles represent the period of 4-h ischemia. Red rectangles represent the period of 12-h reperfusion. Gradient gray rectangles represent no I/R insult. B Protein expression in six treatment groups was assessed by Western blotting. β-actin served as an internal equal loading control. The ratios of C p-JAK2/JAK2, D p-STAT1/STAT1, and E p-STAT3/STAT3 were calculated to determine the activation of JAK/STAT pathway. The relative protein levels of F Bcl-2, G Bcl-xL, and H Bax were also quantified and present in bar graphs. Error bars represent the means ± SD (n = 4). Student’s t-test for comparison between two groups, and one-way ANOVA followed by the Tukey–Kramer post hoc test for comparison among multiple groups were used. *p < 0.05 vs. the value in PBS group after I/R insult, ^&p < 0.05 vs. the value in PRP pre-treatment group after I/R insult. In each bargraph, the white bars from left to right represented pretreatment with PBS, PRP or AG490 without I/R insult, respectively; the black bar represented pretreatment with PBS with I/R insult; the light grey bar represented pretreatment with PRP with I/R insult; the dark grey bar represented pretreatment with AG490 with I/R insult