Glycolytic enzymes dynamically associate with actomyosin during axonal retraction. (A) Western blot showing co-immunoprecipitation (co-IP) of NMII and the glycolytic enzymes GAPDH, PGK, and PK from protein extracts of rat cortical neuronal cultures at DIV3. IgGs are marked by *. (B) Fluorescence images of PLA (in green) and phalloidin staining (in magenta) of the couples of proteins: NMII (phosphorylated RLC) and actin; GAPDH and actin; NMII (heavy chain) and GAPDH; NMII (heavy chain) and PGK; and GAPDH and tubulin. (C) Fluorescence image example with two PLA NMII-PGK punctae and quantification of the position relative to the tip of the growth cone (GC) of 56 PLA NMII-PGK in 26 isolated GCs of calpeptin-treated neurons. (D) Boxplot of PLA signal intensity of neurites of hippocampal neurons treated for 5 min with DMSO (control) or 0.2 U/ml calpeptin of two independent experiments. Statistics: Student’s t test; ***P < 0.001.