Figure 3.

Induction of the CALCA gene by Aza-dC in Rat2 cells. (A) Schematic of the CALCA gene showing the three common exons 1–3 shared by calcitonin (CT) and calcitonin gene–related peptide (CGRP), CT-specific exon 4, and CGRP-specific exons 5, 6. Locations of primers used to measure CT+CGRP (exons 2 and 3), CT (exons 2 and 4), and CGRP (exons 3 and 5) mRNAs are shown. Note that exon 4 is removed from CGRP mRNA. (B) Rat2 cells were treated with 1 or 10 μg/ml Aza-dC (Aza) for six days, or 10 nM trichostatin A (TSA) for one day.

Dimethylsulfoxide (DMSO) was added as a vehicle control (Veh) for the appropriate number of days. RNA from CA77 cells (CA) was used as a positive control. As a polymerase chain reaction (PCR) control, H₂O was added in place of cDNA. Amplification of 18S rRNA served as a control for total RNA levels. (C) NCIH460 cells were treated with 10 μg/ml Aza-dC for five days, then with 10 nM TSA for 1 day (Aza/TSA) prior to harvest. DMSO was added as a vehicle control (Veh). RNA from TT cells was a positive control. As PCR controls, H₂O was added in place of cDNA and β-actin RNA was amplified from all samples.