Figure 9. Induction of hLdhc transcription. (A) Reverse transcriptase polymerase chain reaction (RT-PCR) results showing that hLdhc transcription is induced in nonexpressing cells A375P and C8161 by methylation inhibitor 5'Aza-dC treatment and enhanced by coupled histone deacetylase inhibitor trichostatin A (TSA) treatment. Total RNA was extracted from cells and subjected to RT-PCR to examine the transcription levels in different treatment groups. Multiple transcripts are of splicing variants of the gene. Lane 1: Melanoma C8161 cells with no addition; lanes 2 and 3: C8161 cells treated with 5 µM 5'Aza-dC and with 5 µM 5'Aza-dC + 300 nM TSA, respectively; lane 4: melanoma A375P cells with no addition; lane 5: A375P cells treated with 5 µM5'Aza-dC + 300 nM TSA. The lower panel shows the corresponding β-actin control. (B) RT-PCR showing effects on hLdhc transcription by acetylation inhibitor TSA treatment in melanoma C8161 cells. Lanes 1 to 3: Triplicate assays of cells treated with 5 µM 5'Aza-dC; lanes 4 to 6: triplicate assays of cells treated with 5 µM 5'Aza-dC + 300 nM TSA respectively; lane 7 is PCR negative control. The lower panel shows the corresponding β-actin control. (C) Comparison of the transcription level of full-length hLdhc in C8161 and Hela cells treated with 5'Aza-dC or 5'Aza-dC + TSA (A + T).