Fig. 6.

Immunodetection of DRP-1, phospho-DRP-1, mitofilin, or mitofusin in myoferlin-silenced in PDAC cells. a Total protein extract (10 µg) from HPAF-2 or PANC-1 cells were subjected to SDS-PAGE followed by western blot analysis with specific antibodies against myoferlin, DRP-1, phospho-DRP-1 (ser616), MIC60/mitofilin, or Mitofusin-1. GAPDH was used as a loading control. b Colocalization of phospho-DRP-1 (ser616) and mitochondrial 60 kDa nonglycosylated protein in Panc-1 cells 24 h after myoferlin silencing. Total protein extract (10 µg) were subjected to SDS-PAGE followed by western blot analysis with specific antibodies against myoferlin. HSC70 was used as a loading control. c Tetramethyl rhodamine ethyl ester (TMRE) was used to stain mitochondria in living cells. At 48 h post siRNA transfection, cells expressing or not exogenous DRP-1 were seeded in μ-Slides 8-well at low confluence then loaded with TMRE (1 nM). Representative experiments out of three