Figure 5

Reduction in hnRNPD mRNA and protein levels by pyrrolidine dithiocarbamates (PDTC) treatment in oral cancer cells. (A) Equal amount of total cell lysate proteins from PDTC treated and untreated SCC-4 cells were resolved on 10% SDS-PAGE and subjected Western blotting using monoclonal antibodies against hnRNPD or RelA. Western blot for β-actin served as internal control and was used for normalization for equal loading. (B) Total RNA isolated from SCC-4 cells before or after treatment with PDTC was reverse transcribed and subjected to real time PCR using specific hnRNPD primers. Simultaneously, the PCR was also performed using 18S ribosomal RNA specific primers and served as internal control for normalization of hnRNPD transcript. (C) Total RNA isolated from SCC-4 cells with or without treatment with PDTC was reverse transcribed and subjected to real time PCR using specific hnRNPD, Cyclin D1, TNF-α, IL1-β or IL-8 primers. 18S ribosomal RNA served as internal control for normalization. Values are mean ± SD from three independent experiments performed in triplicate. Results were analyzed using a paired two tailed Student’s t-test and values significantly different from untreated SCC-4 cells have marked by (*P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001).