PKC activates ASIC1 in mesenteric endothelial tubes. (A) Representative immunofluorescence images in mesenteric endothelial tubes showing ASIC1a (red) and STIM-1 (green) distribution. Nuclei are stained with Sytox (blue). (B) Representative images showing lack of Duolink proximity ligation interaction between ASIC1–STIM1 (no observed red puncta). (C and D) Averaged traces (C) and summary data (D) showing Mn²⁺-quenching of fura-2 fluorescence (%) at 360 nm excitation. MnCl₂ (500 µM) was added under vehicle conditions (0 Ca²⁺) or in the absence or presence of PcTX1 (20 nM) following incubation with CPA (10 µM) to deplete intracellular Ca²⁺ stores. Dotted line represents the absence of CPA (Veh). (E) Summary data showing Mn²⁺-quenching of fura-2 fluorescence (%) under basal conditions (dotted line) or following 5 min ACh (10⁻⁶ M) in the absence or presence of PLA₂ inhibitor, MAF (5 µM), and/or PcTX1 (20 nM). Data in D and E represent biological replicates and were analyzed by one-way ANOVA. Post-hoc pairwise comparisons were analyzed between individual groups using Tukey’s multiple comparisons test. (F) Vasodilation responses to AA (10⁻⁸ to 3 × 10⁻⁵ M) following preconstriction with U-46619 in mesenteric arteries. (G) Summary data showing Mn²⁺-quenching of fura-2 fluorescence (%) following 10 min PMA (10⁻⁵ M) in the absence or presence of PcTX1 (20 nM). Biological replicates were analyzed by unpaired t tests.