Pamcyan1 c1

Plasmid DNA encoding AmCyan fluorescent protein (pAmCyan1-C1) was purchased from Clontech Laboratories Inc., Mountain View, CA, USA. The plasmid was propagated in Subcloning Efficiency™ DH5α E. coli competent cells (Invitrogen, Thermo Fisher Scientific, Darmstadt, Germany), then isolated and purified using Qiagen EndoFree Plasmid Mega Kit (Qiagen, Hildesheim, Germany) as per manufacturer’s protocol. CleanCap^® mCherry mRNA was purchased from Tri-Link BioTechnologies LLC, San Diego, CA, USA.

pAmCyan-HPV16E7wt and HPV16E7 deletion mutant constructs (pAmCyan-HPV16E7∆62-66 and pAmCyan-HPV16E7∆71–75) were generated according to Mileo et al. [11 (link)]. All plasmids were transfected using Lipofectamine2000 (Invitrogen, Carlsbad, CA, USA) according to manufacturer’s instruction. Control cells (Mock) were transfected with pAmCyan1-C1 (Clontech Laboratories Inc., Mountain View, CA, USA) empty vector. For GSN silencing assay, FlexiTube siRNA mix for GSN (#SI02664039 and #SI02664046) and negative control siRNA (#1022076) were purchased from Qiagen (Qiagen Corporation, Gaithersburg, MA, USA). C33A cells were co-transfected with DNA plasmids and siRNAs according to Qiagen instructions. The transfected cells were then cultured for 48 h before processing.