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. 2020 Aug 4;11(4):e01728-20. doi: 10.1128/mBio.01728-20

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Copyright © 2020 Ni et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 6 — PPP6C interacts with STING and dephosphorylates STING. (A and B) HA-tagged STING and FLAG-tagged PPP6C were transfected individually or together into HEK293T cells for 30 h. Cell lysates were immunoprecipitated with anti-FLAG (A) or anti-HA (B) antibodies and then immunoblotted with the indicated antibodies. (C) FLAG-tagged PPP6C was cotransfected with full-length (FL), N-terminal (N; aa 1 to 195), or C-terminal (C; aa 181 to 379) HA-tagged STING into HEK293T cells for 30 h. Cell lysates were immunoprecipitated with anti-HA antibody and then immunoblotted with the indicated antibodies. (D) FLAG-tagged STING was cotransfected with full-length (FL), N-terminal (N; aa 1 to 160), or C-terminal (C; aa 151 to 305) HA-tagged PPP6C into HEK293T cells for 30 h. Cell lysates were immunoprecipitated with anti-HA antibody and then immunoblotted with the indicated antibodies. (E) HEK293T cells were cotransfected as indicated with IFN-β-luc, pRL-TK, cGAS and STING plasmids along with plasmids expressing wild-type PPP6C or the indicated PPP6C mutants. Luciferase activity was measured after 30 h. (F and G) EA.hy926 cells were transfected with NS or PPP6C siRNA for 72 h and then transfected with dsDNA (F) or poly(I:C) (G) for the indicated times. Cell lysates were immunoblotted with the indicated antibodies. The data shown are representative of three independent experiments. Data in panel E are presented as mean + SD. *, P < 0.05 by Student's t test.