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. Author manuscript; available in PMC: 2019 Aug 1.
Published in final edited form as: Mol Neurobiol. 2017 Dec 27;55(8):6215–6226. doi: 10.1007/s12035-017-0836-4

Figure 5. Phosphorylation of SAP102 on Ser632 is important for SAP102 clustering in spines.

Figure 5

a, Hippocampal neurons were co-transfected with FLAG-SAP102/DsRed, FLAG-SAP102ΔI2/DsRed, FLAG-SAP102 S632A/DsRed, or FLAG-SAP102 S632D/DsRed. SAP102ΔI2, SAP102 S632A and SAP102 S632D displayed reduced synaptic targeting. FLAG-SAP102 enrichment in spines was significantly reduced by TBB treatment. Scale bar, 5 μm. b, The mean fluorescence intensity of spines compared with that in adjacent dendrites. n = 10–15 neurons from three transfections. *, p < 0.05, t test with Bonferroni’s correction after ANOVA. c, Distribution of proteins in synaptic and extrasynaptic membrane fractions. Equal amounts of total proteins were loaded from each of the fractions and probed with SAP102 Ser632 phosphorylation state-specific antibody, pan-SAP102 antibody, and the early endosomal marker EEA1 antibody. d, Ratio of synaptic to extrasynaptic membrane fractions of pSAP102 and pan-SAP102. The experiment was repeated three times and quantified using ImageQuant LAS TL software. Data represent means ± S.E. (*, p < 0.05, Student’s t test).