C. trachomatis infection increases miR-30c abundance in multiple cell types. (A) Heat map represents log2 fold changes of several miRNAs derived from RNA sequencing. miRNAs reported to be pro-apoptotic are labeled in green, and those reported to be anti-apoptotic are labeled in red. (B) Graph represents the log2 fold changes of miR-30c determined by miRNA deep sequencing of HUVECs after 12 and 24 h of C. trachomatis (C.tr) infection compared with noninfected samples. (C) Graph represents quantification of miR-30c up-regulation upon C. trachomatis infection by qRT-PCR. U6 snRNA was used as endogenous control for qRT-PCR. Cells were infected for 12, 24, and 36 h, and fold changes were normalized to miR-30c expression of noninfected cells at 36 h. Fold change with qRT-PCR for HUVECs (± SD) at 24 h, 2.96 ± 0.47; and 36 h, 4.87 ± 0.46 (n = 6). (D and E)Northern blots of HUVEC and hFIMB cell RNA, probed for U6 SnRNA, miR-30c, and the chlamydial small RNA, CtrR5. Fold change with Northern blot for HUVECs (± SD) at 24 h, 3.9 ± 0.58; and 36 h, 5.76 ± 0.72 (n = 5). Fold changes were normalized to miR-30c expression in noninfected cells at 12, 12, and 36 h, respectively (n = 6 experiments). All data represents mean ± SD. Asterisks denote significance by Student’s t test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, nonsignificant. See also Fig. S1.