Figure 2
SH-SY5Y differentiation is driven by an autocrine secretory signal in CHC22 knockdown cells. (A) Schematic of experimental procedure collecting the media from cells with reduced CHC22 expression and adding it to fresh cells sequentially over 3 days. (B) Phase contrast images of cells spiked with the concentrated, secreted proteins from CHC22 knocked down cells and controls. Neurites quantified have been coloured in red to improve visualisation of the effect. (C) and (D) Quantification of the Neurite length per cell and percentage of cells bearing neurites (greater than the length of two cell bodies) respectively according to methodology described in Nahorski et al.7. Results represent at least three repeats. Statistics represent one way ANOVA (P < 0.001) with Bonferroni’s multiple comparison post hoc test (on Arcsine transformed data for percentages) (p < 0.05, p < 0.01, p < 0.001 are *, **, *** Respectively). (E) Western blot demonstrating the knockdown in cells from which secretions were collected and the CHC22 levels in cells differentiating in response to addition of these secretions.
