MINK1 deficiency impairs ERK, p38, Akt, MEK1/2, and MKK3/MKK6 phosphorylation. (Ai,Ci,Ei,Gi,Ii) Immunoblot analysis of WT and MINK1^−/− platelets, stimulated with collagen (0.8 μg/mL) for 3 minutes in the absence or presence of apyrase (1 U/mL) or tirofiban (2 μg/mL), with antibodies recognizing phosphorylated ERK1/2 Thr²⁰²/Tyr²⁰⁴ (T²⁰²/Y²⁰⁴), total ERK1/2, phosphorylated p38 Thr¹⁸⁰/Tyr¹⁸² (T¹⁸⁰/Y¹⁸²), total p38, phosphorylated Akt Thr³⁰⁸, phosphorylated Akt Ser⁴⁷³, total Akt, phosphorylated MEK1/2 Ser^183/221, total MEK1/2, phosphorylated MKK3 Ser¹⁸⁹/MKK6 Ser²⁰⁷, and total MKK3. (Aii-iii,Cii-iii,Eii-iii,Gii-iii,Iii-iii) Results were quantified and presented as mean ± SEM from at least 3 independent experiments (*P < .05, **P < .01, ***P < .001; paired Student t test). (Bi,Di,Fi,Hi,Ji) Immunoblot analysis of WT and MINK1^−/− platelets, stimulated with thrombin (0.025 U/mL) for 3 minutes in the absence or presence of apyrase (1 U/mL) or tirofiban (2 μg/mL), with antibodies recognizing phosphorylated ERK1/2 Thr²⁰²/Tyr²⁰⁴ (T²⁰²/Y²⁰⁴), total ERK1/2, phosphorylated p38 Thr¹⁸⁰/Tyr¹⁸² (T¹⁸⁰/Y¹⁸²), total p38, phosphorylated Akt Thr³⁰⁸, phosphorylated Akt Ser⁴⁷³, total Akt, phosphorylated MEK1/2 Ser^183/221, total MEK1/2, phosphorylated MKK3 Ser¹⁸⁹/MKK6 Ser²⁰⁷, and total MKK3. (Bii-iii,Dii-iii,Fii-iii,Hii-iii,Jii-iii) Results were quantified and presented as mean ± SEM from at least 3 independent experiments (*P < .05, **P < .01; paired Student t test).