Figure 7: Phosphorylation of SNAP-23 at Ser95 by PKM2 promotes tumour cell exosome release. | Nature Communications

Figure 7: Phosphorylation of SNAP-23 at Ser95 by PKM2 promotes tumour cell exosome release.

From: Pyruvate kinase type M2 promotes tumour cell exosome release via phosphorylating synaptosome-associated protein 23

Figure 7

(a) Constructs of His-tagged RNAi-resistant SNAP-23 (R-SNAP-23) and SNAP-23 (Ser95→Ala95) (R-SNAP-23 (Ser95→Ala95)), in which three nucleotides in siRNA target sequence were mutated without altering amino acid sequence. (b,c) Effect of SNAP-23 phosphorylation at Ser95 on PKM2-mediated release of exosomes from A549 cells. A549 cells were transfected with PKM2 siRNA and His-tagged plasmids expressing SNAP-23, R-SNAP-23, R-SNAP-23 (Ser95→Ala95), SNAP-23 (Ser20→Glu20) or SNAP-23 (Ser95→Glu95). The cellular protein levels of SNAP-23, PKM2 and His tag and the exosome concentration in cell culture medium were assayed by WB and NTA after 24 h culture, respectively. Data are presented as the mean±s.e.m. of three independent experiments. *P<0.05. **P<0.01. ***P<0.001 as determined by the one-way ANOVA test.

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