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. 2019 Oct 30;476(20):3081–3107. doi: 10.1042/BCJ20190608

Figure 8. Phosphorylation of STX12 at Ser139 by SGK3 enhances the association with the SNARE complex.

Figure 8.

(A) The indicated knock-out (KO), and KO cells stably re-expressing wild-type or STX[S139A] HEK293 cells were serum-starved overnight, treated for 1 h with 14H (1 µM) or with DMSO, followed by stimulation with IGF1 (50 ng/ml) for 15 min. GFP-STX12 WT and GFP-STX12 S139A were immunoprecipitated using anti-GFP beads and the immunoprecipitates were subjected to western blot analysis using the indicated antibodies. STX12 phosphorylation was analyzed by Phos-tag assay (° phosphorylated, • non-phosphorylated). Control immunoblots were performed on normal gels with the indicated antibodies. Immunoblots were developed using the LI-COR Odyssey CLx Western Blot imaging system analysis with the indicated antibodies at 0.5–1 µg/ml concentration. Quantification of the respective binding is also provided.