Fig. 8: Design and validation of A312V transgenic mice in wound infection models.

a–e The A312V mutation in mP2X7 does not alter its primary function. No significant difference in current density (pA/pF) was observed between ^m7WT and ^m7A312V (a). Concentration-response curves to ATP of ^m7WT and ^m7A312V indicate similar ATP affinities (b; solid lines were fits of the Hill 1 equation, n = 5). Representative currents demonstrate that current facilitation is inherent to mP2X7 (c), with no significant difference observed between ^m7WT and ^m7A312V (d). Compared to the sustained ATP stimulation-induced current (I_max), the initial ATP-induced current (I₁) is typically smaller (c). Macropore permeability of P2X7 between ^m7WT and ^m7A312V remains comparable (e). Sample sizes (n) of a, d and e are annotated in the plots. Statistical significances were assessed using two-tailed unpaired t-test. f Flowchart depicting the full-thickness wound infection model. g Photographs of wounds treated with PSFL1191 (7.5 mg/ml, 50 μL, subcutaneous) and solutions (control) for 0, 1, 3, 5, and 7 days (scale bar = 2 mm). The green dashed circle outlines the wound’s edge. h Changes in wound size following various treatments. Significant difference between P2rx7^A312V/A312V-PSFL1191 and P2rx7^+/+-PSFL1191 treatments was assessed using two-way ANOVA followed by Tukey’s multiple comparisons test, F (21, 294) = 5.442. i Wound area of P2rx7^A312V/A312V and P2rx7^+/+ after continuous administration for 7 days (Statistical significances were assessed using two-way ANOVA followed by Tukey’s multiple comparisons test, F (21, 294) = 5.442). Sample sizes are annotated under the boxes. Boxes and lines in the boxes represent inter-quantile range (Q3–Q1) and median of relative area, respectively. Lower and upper whiskers show the minimum and maximum values of the relative area. All summary data are expressed as mean ± SEM from independent cells (a-e) or animals (h and i).