Anti actb a2228

cDNAs for human XPC and TDG were generated from total RNAs isolated from human NTERA-2 (NT2) cells. Mammalian expression plasmids were cloned using the pHAGE-EF1α-STEMCCA construct (Sommer et al. 2009 (link)), in which OCT4, KLF4, SOX2, and c-MYC were replaced with cDNAs for XPC, RAD23B, RAD23B–CETN2, or mCherry, which was used as a control (pHAGE-EF1α-XPC, pHAGE-EF1α-RAD23B, pHAGE-EF1α-RAD23B–CETN2, and pHAGE-EF1α-mCherry, respectively). For expressing full-length and truncated human TDG in Escherichia coli, N-terminal His6-tagged TDG was cloned into a pST44 polycistronic expression plasmid (Tan et al. 2005 (link)). Constructs for XPC expression in Sf9s were described previously (Fong et al. 2011 (link)). The TDG-HaloTag construct for single-molecule imaging experiments was subcloned into the pTRE3G plasmid (Clontech). shRNAs against XPC were cloned using the pLKO.1 system as described previously (Moffat et al. 2006 (link)); sequences are provided in Supplemental Table S2. The commercial antibodies used were as follows: anti-ACTB (A2228) from Sigma-Aldrich, anti-XPC (A301-122A) and anti-RAD23B (A302-306A) from Bethyl Laboratories, anti-CETN2 (15977-1-AP) from ProteinTech, anti-5mC (33D3) from Diagenode, and anti-SSEA4 (clone MC-813-70) and anti-TRA-1-60 (clone TRA-1-60-R) from Biolegends.