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. Author manuscript; available in PMC: 2021 Feb 24.
Published in final edited form as: Nat Cell Biol. 2020 Aug 24;22(9):1064–1075. doi: 10.1038/s41556-020-0562-4

Figure 2 |. PD-L1 is deacetylated predominantly by HDAC2.

Figure 2 |

a. IB analysis of WCL and GST pull-down products derived from 293T cells transfected with Myc-p300, GST-PD-L1 C-tail in the presence or absence of the SIRT inhibitor, 5 mM NIA, or the HDAC inhibitor, 1 μM TSA, overnight. b. IB analysis of WCL and anti-HA IPs derived from 293T cells transfected with Myc-p300, HA-ins-PD-L1 and/or indicated Flag-tagged deacetylases. c. IB analysis of WCL and anti-HA IPs derived from 293T cells transfected with indicated constructs to examine PD-L1 acetylation levels. d. IB analysis of WCL and anti-Myc IPs derived from 293T cells transfected with HA-p300, Myc-PD-L1 and/or Flag-HDAC2. e. IB analysis of WCL and anti-PD-L1 IPs derived from MDA-MB-231 cells transfected with control siRNA or siRNAs targeting indicated HDACs. f. IB analysis of WCL and anti-PD-L1 IPs derived from MDA-MB-231 cells transduced with shRNAs against HDAC2 or GFP. g. IB analysis of WCL and anti-PD-L1 IPs derived from MDA-MB-231 wild-type (WT) or HDAC2 KO cells. h. IB analysis of WCL and anti-PD-L1 IPs derived from MDA-MB-231 cells treated with the indicated HDAC inhibitors: Santacruzamate A (SCA), 20 μM; ACY1215, 40 μM; ACY957, 20 μM, for 3 hrs. i. IB analysis of WCL and anti-PD-L1 IPs derived from MDA-MB-231 WT or HDAC2 KO cells, treated with or without 50 μM HDAC2 inhibitor (HDAC2i) for 4 hrs.

The western blots in a-i were performed for n=2 independent experiments with similar results. Unprocessed immunoblots are shown in Source Data Fig. 2.