Figure S3.

Calcium increased the asymmetry of armc4^−/− sperm model flagella.

(A) Traces of armc4^−/− sperm reactivated with ATP + ADP under EGTA or pCa4 conditions with or without 10 μM sodium orthovanadate. ATP + ADP induced highly curved flagella, which were relaxed by the addition of orthovanadate. Scale bar, 10 μm. (B) Quantification of armc4^−/− sperm flagellar curvature. For immotile or irregularly vibrating sperm, one frame per flagellum was traced. Under both EGTA and pCa4 conditions, ATP + ADP induced higher curvature, and dynein ATPase inhibitor orthovanadate significantly reduced the curvature. The pCa4 condition led to higher curvature compared with the EGTA condition when activated by ATP + ADP. All data are shown with a mean (bar graphs) + SE (error bars). (C) Classification of armc4^−/− sperm model motility. Most of the armc4^−/− sperm models showed irregular, vibrating movement or were immotile. Orthovanadate (EGTA + V and pCa4 + V) completely stopped their motility. (B, C) All data were collected from n = 9 (armc4^−/− EGTA), 6 (armc4^−/− EGTA + vanadate), 13 (armc4^−/− pCa4), and 6 (armc4^−/− pCa4 + vanadate) sperm flagella.