pCSPG4 protein expression in pancreatic tissues.

Immunoblotting confirmed the specificity of polyclonal H-300 (A) and monoclonal LHM2 antibodies (B), generated against recombinant protein and melanoma-derived antigens, respectively. The images demonstrate differences in size between pancreatic (band-2) and melanoma (SK-MEL-28, band-1) antigens, but not cervical carcinoma (HeLa) antigens (C), and reveal the frequent existence of oversized isoforms in SCA and PDAC (band 3, C–D). GAPDH was used as the loading control. HiMark (Invitrogen) was used as the molecular weight marker. (E) Western blot and (F) FACS analyses of CSPG4 siRNA-transfected Panc1 cells further confirmed the specificity of the used antibodies and established the knockdown efficacy of the two siRNA sets (si1 and si2) at approximately 75% after 48 h post transfection, compared to the negative control siRNA (neg.si). QRT-PCR confirmed the efficacy of the knockdowns at the mRNA level.