ANO5 does not require “scrambling domain” to rescue defective repair. (A) Deconvolved images of FM1-43 infiltration in ANO5-KO, ANO5-KO + hANO5, or ANO5-KO + ANO5-1-5 fibers before and following injury. Scale bars = 10 µm. (B and C) FM1-43 accumulation time course (B) and AUC values (C) for individual ANO5-KO, KO + hANO5, KO + ANO5-1-5, or WT fibers. ANO5-KO n = 11 fibers from three mice, ANO5-KO + hANO5 n = 16 fibers from three mice, ANO5-KO + ANO5-1-5 n = 10 fibers from three mice. Blue dotted line represents the average FM1-43 F/F_maxKO value from 11 injured WT fibers. (D) Representative images of Ca²⁺ transients visualized in damaged fibers with Calbryte 520. (E and F) Calbryte 520 time course (C) and AUC values (F) for individual ANO5-KO, KO + hANO5, KO + ANO5-1-5, or WT fibers. ANO5-KO n = 15 fibers from three mice, KO + ANO5 n = 16 fibers from two mice, KO + ANO5-1-5 n = 13 fibers from two mice, WT n = 9 fibers from three mice. (G and H) ANXA2 F/F₀ time course (G) and AUC values (H) for individual ANO5-KO fibers + ANO5 or ANO5-1-5. KO + ANO5 n = 16 fibers from two mice, KO + ANO5-1-5 n = 16 fibers from two mice. Blue and red dotted lines represent ANXA2 accumulation in WT and ANO5-KO fibers, respectively. (I and J) ANXA6 F/F₀ time course (I) and AUC values (J) for individual ANO5-KO fibers + ANO5 or ANO5-1-5. KO + ANO5 n = 24 fibers from five mice, KO + ANO5-1-5 n = 18 fibers from three mice. Blue and red dotted lines represent ANXA6 accumulation in WT and ANO5-KO fibers, respectively. Data are mean ± SEM. *, P < 0.05. Scale bar = 10 μm.