In vivo miR-148a inhibition diminished calcium mobilization via GPVI and FcγRIIA stimulation. A total of 1 × 106 platelets from anti–miR-148a– or control-treated mice were labeled with Fluo-4-AM. Baseline intracellular calcium was assessed for 60 seconds, and indicated doses of agonists were added to the platelets: (Ai-Aii) 1 µg/mL IV.3 + 4 µg/mL GAM and 2 µg/mL IV.3 + 8 µg/mL GAM were added, respectively, at the indicated time. (Aiii) Quantification of peak calcium fold change (n = 3). (Aiv) Quantification of area under the curve (AUC) for calcium mobilization curve (n = 3). AUC was calculated in Prism software. (Bi-Biii) 2.5 µg/mL CRP, 6.25 µg/mL CRP, and 12.5 µg/mL CRP were used to induce calcium influx, respectively. (Biv-Bv) Quantification of peak calcium fold change and AUC were performed as discussed above (n = 3).
