Fig. 7: PPP1CA–SETX dephosphorylates histone H3T11 to regulate autophagy in mammals. | Cell Discovery

Fig. 7: PPP1CA–SETX dephosphorylates histone H3T11 to regulate autophagy in mammals.

From: Glc7/PP1 dephosphorylates histone H3T11 to regulate autophagy and telomere silencing in response to nutrient availability

Fig. 7

a Immunoblot analysis of H3pT11 in HeLa cells transfected with plasmids overexpressing PPP1CA, PPP1CB and PPP1CC. b Immunoblot analysis of H3pT11 in control (shCtrl) and PPP1CA-knockdown (shPPP1CA#1, shPPP1CA#2) HeLa cells. c The purified recombinant PPP1CA dephosphorylates H3T11 as determined by in vitro dephosphorylation assay. d Analysis of autophagy activity in control (shCtrl) and PPP1CA-knockdown (shPPP1CA#1, shPPP1CA#2) HeLa cells. e ChIP-qPCR analysis of PPP1CA occupancy at autophagy-related genes (ULK1, ATG7, ATG12, LC3) in HeLa cells. f ChIP-qPCR analysis of H3pT11/H3 occupancy at autophagy-related genes in control (shCtrl) and PPP1CA-knockdown (shPPP1CA) HeLa cells. g RT-qPCR analysis of the transcription of autophagy-related genes in shCtrl and shPPP1CA HeLa cells. h Co-IP and reciprocal IP showing the interaction between PPP1CA and SETX. i ChIP-qPCR analysis of SETX occupancy at autophagy-related genes in HeLa cells. j ChIP-qPCR analysis of PPP1CA occupancy at autophagy-related genes in control (shCtrl) and SETX-knockdown (shSETX) HeLa cells. k ChIP-qPCR analysis of H3pT11/H3 occupancy at autophagy-related genes in shCtrl and shSETX HeLa cells. The telomere-proximal genes, C1S and CDL163L1 were used as controls. l RT-qPCR analysis of the transcription of autophagy-related genes in shCtrl and shSETX HeLa cells. m Analysis of autophagy activity in shCtrl and shSETX HeLa cells. For b, d–g, i–m, data represent the mean ± SE of three biological independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001.

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