Skip to main content
. 2018 Nov 12;13(11):e0207205. doi: 10.1371/journal.pone.0207205

Fig 2. The influence of S226 phosphorylation on the interaction between SNX5 and SNX1 or SNX2.

Fig 2

(A) FLAG pull-down assay was performed on lysates of 8505C cells which were expressed SNX5-FLAG wild type (Wt), or a mutant, T139A/S141A/S142A (TSS-AAA)), T139E/S141E/S142E (TSS-EEE), S151A/S152A/S157A (SSS-AAA), S151E/S152E/S157E (SSS-EEE), S226A, S226E, by an adenovirus expression system. FLAG fused proteins (SNX5), SNX1 and SNX2 were detected by immunoblotting with anti-FLAG, anti-SNX1, anti-SNX2 antibodies. (B) Immunoprecipitation using anti-SNX1 antibody was performed on lysates of 8505C cells which were expressed FLAG-tagged SNX5 wild type, S226A, and S226E by adenovirus expression system. FLAG fused proteins and SNX1 were detected by immunoblotting with anti-FLAG and anti-SNX1 antibodies, respectively. (C) The band intensity of (B) were measured and quantified. (D) CD spectra of MBP fused SNX5 wild type (blue), S226A (red), S226E (green). (E) Immunocytochemical staining of 8505C cells which were expressed by SNX5-FLAG wild type, S226A, and S226E. The cells were stained with anti-FLAG (green), anti-SNX1 (red) and anti-SNX2 (red) antibodies and observed using confocal microscopy. Scale bars: 10 μm.