Hsp27 Knockdown Cell Generation

Hsp27 knockdown (Hsp27-KD) cells were generated using Hsp27-specific short hairpin RNA (shRNA) (National RNAi Core Facility, Academia Sinica, Taiwan) as previously described [20 (link), 38 (link)–41 (link)]. The target sequence for the human Hsp27 mRNA (NM_001540) was 5′-CCGATGAGACTGCCGCCAAGT-3′. The MISSION non-target shRNA control vector (SHC002) was used as a scrambled control (Sigma Chemical Co.) [38 (link)–41 (link)]. HCC cells were transfected with the knockdown vector or the parental vector (pLKO.1<-puro) and then selected with puromycin. The transfection and generation of stably transfected cell lines were performed as previously described [40 (link)–42 (link)].