S135F mutant decreases CerS1 localization to mitochondria. A) CerS1 localizes to mitochondria in the presence of WT Hsp27 more than S135F in HT-22 cells. Quantification of super-resolution SIM colocalization of FLAG (CerS1) and MitoTracker Deep Red (mitochondrial marker) used PCC and MOC (*P < 0.05 for PCC, *P < 0.05 for MOC, n = 31 and n = 35). B) SIM quantification of FLAG and calnexin (ER marker) colocalization (P = 0.797 for PCC, P = 0.188 for MOC, n = 24 and n = 27). C) CerS1 localizes to the ER in the presence of WT and S135F Hsp27 in HT-22 cells. SIM quantification of FLAG and calnexin colocalization using PCC and MOC (P = 0.131 for PCC, P = 0.70 for MOC, n = 15 and n = 18). D) SIM quantification of FLAG and calnexin colocalization was quantified using PCC and MOC (P = 0.324 for PCC, P = 0.287 for MOC, n = 9 and n = 10). E) SIM quantification of calnexin and MitoTracker Deep Red (P = 0.324 for PCC, P = 0.287 for MOC, n = 9 and n = 10).