Fig. 5

SARS-CoV-2 N Protein induces expression of HDAC6. A Cellular lysates were prepared from A549 cells either transfected with increasing concentration of pcDNA3.1-N (1–3 μg) or transfected with empty pcDNA3.1 vector. Expression of HDAC6, N protein and GAPDH was analysed via western blotting. B Co-immunoprecipitation analysis was performed in A549 cells either pcDNA3.1-N protein-transfected or empty vector-transfected A549 cells using anti-G3BP1 antibody and immunoblot analysis was performed using HDAC6 and N protein-specific antibodies. C Total RNA was isolated from A549 cells either transfected with empty vector or pcDNA3.1-N protein followed by qRT-PCR using hdac6 and gapdh-specific primers. ns = not significant, unpaired t-test. D A549 cells were either transfected with empty vector or pcDNA3.1 N protein followed by CHX treatment at 18 h post-transfection. Cells were harvested at indicated time post-CHX treatment (0, 6, 12, 18) and HDAC6, SARS-CoV-2 N protein and GAPDH expressions were analysed via immunoblotting. E A549 cells were either transfected with pcDNA3.1-N protein or empty vector followed by immunoprecipitation with anti-K48-ubiquitin antibody. Immunoblot analysis was performed with anti-HDAC6 antibody. F SARS-CoV-2-infected (24–32 hpi) or mock-infected lysates were pulled down using with anti-K48-ubiquitin antibody. Immunoblot analysis was performed with anti-HDAC6 antibody