Pcmv c dsred

For the overexpression assay and the localization assay, expression vector and fluorescence-labeled vector were constructed. In brief, the PLIN5/ATF1/ATF4 CDS region was amplified by the cDNA library of HepG2 cells using KOD-Plus-Neo DNA polymerase (#KOD-401, TOYOBO, Shanghai, China). After gel extraction, the PLIN5 CDS fragment was cloned into the digested pcDNA3.1 vector (digestion sites, HindIII and BamHI) using a seamless cloning kit (#C112-01, ClonExpress II One Step Cloning Kit, Vazyme, Nanjing, China). For the localization assay, the gene CDS region was cloned into the digested pCMV-C-Dsred (#D2624, Beyotime Biotechnology, Nanjing, China) or pCMV-C-EGFP (#D2626, Beyotime Biotechnology, Nanjing, China). For the luciferase reporter assay, the promoter region of PLIN5 (about 2000 bp upstream of the transcription initiation site of PLIN5) was cloned into the digested pGL3-basic vector (digestion sites, KpnI and XhoI).

Trans 5α was purchased from Transgen Biotech (Transgen, Beijing, China). Plasmids including pCMV-C-EGFP (#D2626), pCMV-C-DsRed (#D2624) and pCMV-N-FLAG (#D2722) were purchased from Beyotime (Beyotime, Nanjing, China).