Figure 2

5AZA and TSA up-regulate the expression of OX40L and 41BBL mRNA in SW620 cells. (A) OX40L, and (B) 41BBL mRNA was quantified in SW620 cells using qPCR. Cells were plated and treated with 5AZA-dC (20 uM), TSA (250 nM) or 10Gy radiation (IR). Adherent cell were collected after 48 (gray bar) and 72 hr (black bar) and mRNA values were compared to the level of gene expression see in untreated control samples (white bar), which was set to 1. Values represent mean ± SEM of technical replicates. Experiments were repeated at least three times with similar results. (C) Cells were plated and untreated (DMSO) or treated with 5AZA-dC (20 uM), TSA (500 nM, 250 nM or 125 nM) or 10Gy radiation (IR). Both floating and adherent cells were collected after 48 h of treatment and tumor cell viability was determining using trypan blue dye exclusion. Values represent mean ± SEM of three independent experiments. (D) OX40L and (E) 41BBL mRNA level was quantified after 8 (gray bar) and 24 hr (black bar) and compared to the level of gene expression in untreated control samples (white bar). Values represent mean ± SEM of technical replicates. Experiments were repeated at least three times with similar results. (F) Cells were plated and left untreated (DMSO) or treated with 5AZA-dC (20 uM), TSA (500 nM, 250 nM or 125 nM) or 10Gy radiation (IR). Adherent cells were collected after 24, 48, and 72 h of treatment and live tumor cell number was determining using trypan blue dye exclusion. Values represent mean ± SEM of three independent experiments. Significant P-value shown in the indicated groups was determined at 48 h. *indicates P value of <0.05, **indicates P value of <0.001.