FIGURE 2.
Effect of SPINK3 on sperm membrane potential (Em) and associated pathways. (A) Em measurements of DISC3(5)-loaded sperm, incubated for 60 min in non-capacitating (NC, gray bar) or capacitating (CAP, black bars) conditions containing the specified SPINK3 concentrations. Data represent mean ± S.E., n = 4, ** values significantly different (p < 0.01) compared to CAP; (B,C) after sperm incubations as detailed in (A), western blot analyses were performed with anti pSrcY416 antibodies (B) (clone D49G4), further stripped and re-probed with anti-tubulin (α-Tub) antibodies. Alternatively, western blot analyses were performed with anti-pPKAs (clone 100G7E) (C), further stripped and re-probed with anti-pTyr (α-pY) (clone 4G10) (D). Western blot analyses are representative of experiments repeated at least three times. p-hexokinase (116 kDa, α-pHK) detected with anti-pY, is showed as loading control.