Gfp lc3b

The human HTT-exon1 constructs (Q72 and Q25) were synthesized by Thermo Fisher Scientific and subcloned into the pcDNA/DEST40 vector (Thermo Fisher Scientific, #12274-015). The mouse full-length Hipk3 cDNA was codon-optimized and synthesized by Thermo Fisher Scientific and subcloned into the pTT5SH8Q2 vector (NRC Biotechnology) with N-terminal GFP tag. The human or rat Hipk3 cDNA failed to express efficiently in STHdh cells. The Q72-GFP construct (expressing Methionine-Q72-GFP) was cloned by PCR amplification from the HTT-exon1 constructs, and ligation to the pcDNA-GFP plasmid. The QuikChange Site-Directed Mutagenesis Kit (Agilent Technologies) was used to generate the point mutation constructs: kinase-dead mouse Hipk3 (K226M and D322N), DN-MAPK11 (TGY (180-182) to AGF). The MAPK11 (#20355) and GFP-LC3B (#24987) construct was obtained from Addgene.

SARS-CoV-2 ORF3a (Gene ID: 43740569) with HA-Tag was cloned into the pCAGGS through restriction sites EcoRI and BamHI. Plasmids GFP-mCherry-LC3B (#123230) and GFP-LC3B (#11546) were purchased from Addgene (Watertown, MA, USA). Transfection reagent (Yeasen, Shanghai, China) was used according to the manufacturer’s protocol.