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. 2015 Aug 31;603:77–83. doi: 10.1016/j.neulet.2015.07.019

Fig. 3.

Fig. 3

ERK1/ERK2 activation by CaSR contributes to neurite growth.

(A) Images of non-transfected E18 SCG neurons double labelled for phospho-ERK1/ ERK2 and βIII-tubulin after 24 h with and without 10 ng/ml NGF in media containing either 0.7 mM or 2.3 mM [Ca2+]o. (B) Images of double labelled E18 SCG neurons after 24 h incubation in NGF-free medium containing 2.3 mM [Ca2+]o after transfection with either pcDNA3.1 or pFLCaSR. Scale bar = 20 μm. (C and D) Quantification of phospho-ERK1/ERK2 immunofluorescence (C) and total ERK1/ERK2 immunofluorescence (D) in E18 SCG neurons cultured for 24 h in NGF-free medium containing 2.3 mM [Ca2+]o plus 50 μM Boc-D-FMK transfected with either pFLCaSR or an pcDNA3.1. Mean ± sem of data from 40 neurons per condition. ***P < 0.001, unpaired t-test with Welch’s correction. Total length (E) and branch point number (F) of E18 SCG neurons cultured without NGF for 24 h in media containing 2.3 mM [Ca2+]o plus 50 μM Boc-D-FMK and either 10 μM U0124 or 10 μM U0126 after transfection with either pFLCaSR or pcDNA3.1. Mean ± sem of data from 214 to 249 neurons per condition. ****P < 0.0001, ANOVA with Bonferroni’s post-hoc test.