Fig. 1

γ-Enolase associates with the intracellular domain of Trk in SH-SY5Y cells. A Representative co-immunoprecipitation (top) and quantification (bottom; as relative amount of γ-enolase in the immunoprecipitate in wild-type vs. transfected cells) of γ-enolase and Trk from wild-type and transfected SH-SY5Y cells with down-regulated γ1-syntrophin expression (Stealth RNAi SNTG1). Wild-type and transfected cells were differentiated by serum deprivation for 24 h, and then lysed and immunoprecipitated (IP) with either non-specific rabbit IgG or rabbit anti-pan-Trk antibodies (Trk). The immunoprecipitates (left panel) and lysates for immunoprecipitation (right panel) were analysed by Western blotting probed with a mouse anti-γ-enolase antibody. In case of lysates, β-actin was used as loading control. Data are means ± SD of two independent experiments (n = 2) (one-way ANOVA, Dunnett’s test, *P < 0.05) B Representative images (left) and quantification (right) of double immunofluorescence staining for γ-enolase (green fluorescence) and pan-Trk (red fluorescence) in wild-type and transfected SH-SY5Y cells 24 h after serum deprivation, showing stronger co-localization of γ-enolase with Trk (solid arrows) in wild type cells compared to transfected cells (dashed arrows). Data are means ± SD of the pixels in the third quadrant of scatter plot (cell numbers ≥ 10) (two tailed t test, * P ˂ 0.05). Scale bars: 10 µm