Exogenous ANO5 expression rescues Ano5^−/− MPC Ca²⁺-PLS. (a) Simultaneous whole-cell patch clamp and annexin V–Alexa Fluor 568 binding as an indicator of Ca²⁺-PLS of primary MPCs differentiated for ∼24 h. Images on the left show bright-field micrographs of patched MPCs. Images at the center and right show annexin V binding. Scale bar, 10 μm. (b) Quantification of Ca²⁺-PLS in MPCs. Statistical significance was measured using a Fisher exact test (*, P = 0.024). (c) Representative traces of ionic currents from MPCs isolated from the same mouse with or without ANO5 virus rescue following 10 min of whole-cell patch clamp with 200 µM Ca²⁺ intracellular solution. > denotes 0 nA. 20mV voltage steps from −60 mV to 140 mV. (d) Quantification of ionic currents from cell lines isolated from two 3-mo-old mice with and without viral rescue (*, P = 0.027, paired t test). (e) Current–voltage relationship ANO5 rescued Ano5^−/− MPCs from panel d.