Fig. 5.

Nefl deletion increases Stat3–stathmin interaction. a Stathmin was immunoprecipitated from sciatic nerve extracts of 34-day-old mice. Western blot shows from left to right, input (Ip) and eluate (E) from wild-type and Nefl−/− mice nerve extracts after anti-IgG control and anti-stathmin pulldown. b Quantification of band intensities in the eluate showed an increased interaction of Stat3 and stathmin in Nefl−/− mice (t = 17.08; P = 0.0034). Bars represent mean ± SEM (one sample t test, n = 3 independent experiments). c Nefl deletion increases Stat3–stathmin interaction in sciatic nerves from pmn mutant mice. d Western blot analyses show similar levels of stathmin in the sciatic nerve extracts of 34-day-old wild-type, pmn and Nefl−/− mice. Histone levels were determined to ensure equal loading of proteins. e Quantification of Stat3 per stathmin levels in the sciatic nerve extracts (input before immunoprecipitation) from 34-day-old wild-type and Nefl−/− mice (n = 3 independent experiments). f Stathmin was immunoprecipitated from NSC-34 cell extracts. Western blot shows from left to right input for IgG, mock, and NFL overexpressing NSC-34 cells. Right blot shows eluate after IgG control pulldown and stathmin pulldown from NSC-34 cells after transfection with mock (only lipofectamine) or, NFL overexpression vector, respectively. g Western blot analysis reveals increased levels of phosphorylated Stat3 (at Y705) in the sciatic nerve extracts of 34-day-old pmn and Nefl−/− in comparison to wild-type mice. Quantification of pStat3 normalized by total Stat3 levels in h pmn mouse compared to wild type (t = 4.928; P = 0.0044; n = 6 independent experiments) and i in Nefl−/− mouse compared to the wild-type mouse (t = 2.865; P = 0.0242; n = 8 independent experiments). Bars represent mean ± SEM