Platelets synthesize the Shh ligand upon agonist stimulation. (A) Amplification chart representing Cq of mRNAs in platelets as indicated. (B) Synthesis of Shh in platelets treated with thrombin (Thr) (0.5 U/mL) for varying time points. PVDF membrane developed for Shh was stripped and reprobed for β-actin. (C) Corresponding densitometric analysis normalized against β-actin. (D-E) Surface externalization of Shh in thrombin (1 U/mL)-induced platelets. (F-G) Shh exposure on EVs released from thrombin (1 U/mL)-stimulated platelets. RP, resting platelets. Data are represented as mean ± standard error of the mean of 3 to 5 independent experiments. *P < .05 as compared with resting platelets; #P < .05 as compared with thrombin-stimulated platelets.