Fig. 1. Ciliobrevin A and ALCAR modulate the axonal retrograde transport of H_CT in motor neurons.

a Speed profiles of H_CT carriers in motor neurons treated with DMSO (black squares) or 50 μM ciliobrevin A (red diamonds) (DMSO: 55 carriers, 10 axons; 50 μM ciliobrevin A: 46 carriers, 8 axons; 3 independent experiments). b, c Effects of 100 μM ciliobrevin A on the accumulation of H_CT in the cell body of ES cell-derived motor neurons. ES cell-derived motor neurons were incubated with AlexaFluor 555-conjugated H_CT and DMSO or 100 μM ciliobrevin A for 2 h. Cells were then acid-washed, fixed, imaged and quantified in C as described below. d, e Effects of 100 μM ciliobrevin A on the accumulation of α-p75^NTR in the soma of ES cell-derived motor neurons. ES cell-derived motor neurons were incubated with α-p75^NTR and DMSO or 100 μM ciliobrevin A for 2 h. Cells were then acid-washed, fixed, permeabilised, stained for α-p75^NTR and quantified in E. f Speed profiles of H_CT carriers in ES cell-derived motor neurons treated with control medium (black squares) or 1 mM ALCAR (green triangles) (control: 53 carriers, 10 axons; 1 mM ALCAR: 61 carriers, 8 axons; 3 independent experiments). g, h Effects of 1 mM ALCAR on accumulation of H_CT (g, h) and α-p75^NTR (i, j) in the cell body of ES cell-derived motor neurons. ES cell-derived motor neurons were treated as described in B-E. The amount of H_CT and α-p75^NTR was quantified as the mean staining intensity per pixel in the cell body. Results in C, E, H, J are expressed as a percentage of the control ± SEM (n = 3 independent experiments; n ≥ 25 cell bodies analysed per condition). *** p < 0.001 (unpaired Student’s t-test)