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Fig. 2. TSA treatment of P19 cells induces the expression of cardiac -actin, Gata4, Bmp4, Mef2c and Nkx2-5. P19 cells were aggregated and treated with 0, 2 nM, 5 nM or 10 nM TSA, or DMSO each day (day 0 to day 4). Total RNA was harvested on days 0, 2, 3, 4 and 6. (I) Northern blot analysis was used to detect the transcripts for cardiac -actin, Gata4, Bmp4 and Brachyury T from 12 µg of total RNA on the days indicated. (II) RT-PCR was performed on the RNA harvested on days 0, 2, 3, 4 and 6, and Southern blot analysis was used to detect for the expression of Nkx2-5 and Mef2c. Lane 17 (+) shows a positive control with RNA from DMSO-induced cardiomyocytes on day 6. RT-PCR controls include the positive control without reverse transcriptase (lane 18), with control RNA (lane 19) and with PCR-H₂O (lane 20). (III, IV) Densitometric quantification was performed for the expression of (III) Gata4 and cardiac -actin by northern blot analysis and (IV) Nxk2-5 and Mef2c by RT-PCR for cells treated with 5 nM TSA. Results are shown as the fold enhancement of transcript levels in TSA-treated cells compared with untreated cells on days 2, 3, 4 and 6. Results are also shown for the fold enhancement of Gata4 and cardiac -actin transcripts on day 6 in cells treated with DMSO compared with untreated cells. Error bars represent the standard error (+ s.e.) of three separate experiments.

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