β actin primers

Silibinin (purity >98%) and puromycin were purchased from Sigma-Aldrich (St. Louis, MO), pifithrin-α was obtained from Calbiochem (Temecula, CA). p53, p21, GADD45α, XPA, XPB, XPC, and XPG antibodies and normal goat serum were from Santa Cruz Biotechnology (Santa Cruz, CA). Horseradish peroxidase (HRP)-labeled anti-thymine dimer antibody was from Kamiya Biomedical Company (Seattle, WA), and antibody to ssDNA (MAB3299) was obtained from Chemicon (Temecula, CA). Genomic DNA and mRNA purification kit, specific primers for human NER factors (XPA, XPB, XPC, XPG, DDB1 and DDB2), p53 and GADD45α as well as PCR-master mix were obtained from Qiagen (Germantown, MD). β-actin primers were obtained from Sigma (St. Louis, MO). IR800 fluorescent dye-labeled anti-mouse IgGs was from LI-COR Biosciences (Lincoln, NE). AlexaFluor 488, 594, 555 and 647 secondary antibodies were purchased from Molecular Probes Inc. (Eugene, OR).

Total RNA was isolated with RNeasy Mini Kit (74106, Qiagen) and measured with nanodrop (NanoDrop™ One, Thermo Scientific, Waltham, MA, USA). IScript™ cDNA Synthesis Kit (1708891, BioRad) was used for cDNA synthesis. Primers for VEGF (Hs00900055_m1), IL6 (Hs00174131_m1), PAI1 (Hs00167155_m1), and p15Ink4b (Hs00793225_m1) were obtained from TaqMan (4331182, Thermo Fisher Scientific), and β-actin primers were obtained from Sigma (05046165001). Quantitative PCR was performed using TaqMan Fast Advanced Master Mix (4444557, Applied Biosystems™, Waltham, MA, USA) and The Applied Biosystems QuantStudio 12K Flex Real-Time PCR System. Further, the following protocol was utilized: denature for 2 min at 95 °C, followed by 40 cycles, each consisting of 95 °C denaturation for 1 s, and 60 °C annealing for 20 s. The relative expression level of the transcripts was analyzed using the 2-△△Ct approach.