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. 2013 Apr 16;8(4):e61991. doi: 10.1371/journal.pone.0061991

Figure 2. Validation of microarray by real-time RT-PCR.

Figure 2

HUVECs transduced with the control vector (Vector, white bars), or the GPR4 expression construct (GPR4, dark bars) were treated with EGM-2/HEM media at pH 8.4, 7.4, or 6.4 for 5 h. Total RNA was isolated and cDNA was synthesized. Real-time RT-PCR quantification of mRNA levels of CXCL2 (A), CCL20 (B), IL8 (C), PTGS2 (D), RELB (E) and TRAF1 (F) was performed. Ct values were normalized to the housekeeping gene GAPDH. The expression level of the target gene in HUVEC/Vector or HUVEC/GPR4 cells at pH 8.4 was set as 1. Error bars indicate the mean ± SEM. *, P<0.05; **, P<0.01; ***, P<0.001; ns, not significant (P>0.05); compared with the pH 8.4 groups. The results shown are the average of at least two biological repeats.