Fig. 3

Myoferlin silencing impacts mitochondrial Ca²⁺ level upon histamine stimulation. A Panc-1 and MiaPaCa-2 cell lines transfected with CMV-Mito-R-GECO-1 plasmid upon histamine stimulation in a Ca²⁺-free medium at different time points (0, 60, 125 s). Histamine injection was done at time point 50 s. Images were acquired with a Nikon A1R microscope. B Quantification of Ca²⁺ level upon histamine stimulation. Fluorescence was monitored over time. Histamine was injected (arrow) at 50 s. Fluorescence was monitored for each individual cell over time (Fn) and normalized to the fluorescence of the first frame of the time lapse (F0). C The peak amplitude was the difference between the normalized fluorescence at 60 and 45 s for the Panc-1 cell line and 65 and 45 s for the MiaPaCa-2 cell line. The total number of cells for each condition was n = 38 (no siRNA), n = 25 (Myof#1 siRNA), n = 33 (Myof#2 siRNA) and n = 41 (Irrelevant siRNA) for the Panc-1 cell line and n = 43 (no siRNA), n = 79 (Myof#1 siRNA), n = 60 (Myof#2 siRNA) and n = 107 (irrelevant siRNA) for the MiaPaCa-2 cell line. Results are presented as mean ± SEM. Tukey’s test was used for statistical analysis. **** p-value < 0.0001. D Western blot validating myoferlin silencing. HSC70 was used as a loading control