Figure 6.

Phosphorylation of the p65 NF-κB subunit at ser536 is required for RANK-mediated inhibition of neurite growth. (a) Representative Western blot of phospho-S536-p65 and total p65, and graph of time course of p65 phosphorylation in cultured SCG neurons following stimulation with 10 ng ml⁻¹ RANKL after 12 h pre-incubation with 10 ng ml⁻¹ NGF. Each data point represents the mean ± s.e.m. of six separate Western blot assays. (b) Sholl profiles and bar charts of (c) total neurite length and (d) branch point number of the neurite arbours of P0 SCG neurons after 24 h incubation in medium containing 10 ng ml⁻¹ NGF following co-transfection with a YFP plasmid together with either an empty control plasmid or plasmids expressing RANK, S536A-p65 or RANK plus S536A-p65. The length and branch point data are expressed as a percentage of data from control-transfected neurons. Means ± s.e.m. of typical experiments are shown (50–90 neurons per condition, **p < 0.001, statistical comparison with the respective controls). Sholl plots from representative experiments are shown. The non-normalized neurite length and branching datasets for all experiments carried out are provided in electronic supplementary material, spreadsheet S6.