Dysregulated calcium homeostasis prevents plasma membrane repair in Anoctamin 5/TMEM16E-deficient patient muscle cells

a Chromatogram for the sequence of the genomic DNA region of patient (middle and bottom) and healthy (top) myoblasts examining the mutation of the ANO5 gene. b Images showing immunostaining and (c) Quantification of the fluorescence intensity of ANO5 immunostaining in healthy (n？=？25 cells) and patient myoblasts (n？=？41 cells, ****p？<？0.0001). Scale bar？=？10？μm. d RT PCR quantification of ANO5 transcript using two independent ANO5 mRNA-specific primers. e Images showing myosin heavy chain 3 (MYH3, Green) and DAPI (nuclear, Red) staining of myoblasts differentiated for 10 days. Scale bar？=？10？μm. f Quantification of the number of myonuclei in healthy and patient myoblasts at indicated days following start of differentiation. g–j Proteins isolated from differentiating myoblasts at indicated days post differentiation are analyzed by Western Blot for differentiation markers. g Representative Blots and h–j Quantification of band intensity for proteins induced during differentiation h Myosin heavy chain 3 (MYH3), i Desmin and j α-actinin normalized to the loading control (β-actin) (n？=？3 independent blots). The β-actin normalized band intensities were not significantly different between healthy and patient myoblasts based on the unpaired multiple t-tests except where indicated by *. (*p？=？0.046 for h and *p？=？0.049 for