Upregulation of TRPCs by ATRA in A549 cells.

<p><b>A,</b> The mRNAs of TRPC1, 3, 4 and 6 were detected in A549 cells by RT-PCR using the primer set in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0067637#pone.0067637.s001" target="_blank">Table S1</a>. The PCR bands for TRPC5 and TRPC7 were negative in A549 cells, but positive in human brain or HepG2. <b>B,</b> The mRNA was detected by real-time PCR in the A549 cells treated with all-<i>trans</i> retinoic acid (ATRA, 1 µM) for 96 hours. The β-actin was used as a housekeeping gene for relative quantification. The 2^(−ΔΔCt) method was used for calculation. <b>C,</b> The TRPC proteins were quantified by Western blotting using anti-TRPC1 (T1E3), anti-TRPC3, anti-TRPC4 (T45E3) and anti-TRPC6 antibodies. Anti-β-actin antibody was used for relative protein quantification (<i>n</i> = 3 independent experiments and each experiment with triplicate samples).