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. 2023 Feb 8;299(3):102990. doi: 10.1016/j.jbc.2023.102990

Figure 4.

Figure 4

Mprocleavage HDAC2 occurs at residues Q261 and Q383.A, HEK-293T cells were transfected with pCDNA3.1-HDAC2-Flag plasmid or its truncated constructs pHDAC2 (190–488)-Flag, pHDAC2 (250–488)-Flag, pHDAC2 (365–488)-Flag. After 30 h of transfection, the cell lysates were analyzed by Western blotting with anti-flag antibody. B, schematic representation of substrate specificity of SARS-CoV-2 Mpro. C, HEK-293T cells were respectively transfected with HDAC2, HDAC2-Q254A, HDAC2-Q261A, HDAC2-Q354A, HDAC2-Q365A, HDAC2-Q381A, and HDAC2-Q383A together with Mpro or empty vector. Western blotting was performed 24 h posttransfection. D, HEK-293T cells were transfected with double mutants Q261A and Q383A of HDAC2 together with Mpro or empty vector. Western blotting was performed 24 h posttransfection. E, schematic representation of HDAC2 with the position of two cleavage sites. F, HEK-293T cells were transfected with a plasmid expressing EGFP-HDAC2 together with Mpro-mCherry or empty vector. After 24 h expressing, images of transfected cells were recorded with GFP and mCherry excitation and emission spectra. Scale bar, 10 μm. HDAC, histone deacetylase; Mpro, main protease; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2.