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. 2016 Dec 7;8(5):7572–7585. doi: 10.18632/oncotarget.13810

Figure 3. Upregulated miRNAs by HDACIs that potentially inhibit CCR6 in CTCL cells.

Figure 3

(A) Heat map and intensity of miRNAs with seed sequence of CCR6 [FC, fold change: > 1.5]. V/D = vorinostat/DMSO, P/D = panobinostat/DMSO. (B) Northern blot analysis of candidate miRNAs with CCR6 seed sequence (miR-96-5p, miR-150, miR-183-5p, miR-194-5p, miR-301b, miR-320c, and miR-371b-5p) showing high fold change (FC: > 1.5) for normal CD4+ cells (CD4-1, and CD4-2), CTCL cell lines, and primary MF samples (n = 2, tumor phase). 5S tRNA (5S), control. (C) Western blot analysis of CCR6 expression in My-La, HH, and HUT78 cell lines transiently transduced with respective candidate miRNAs (Scr: scrambled; 96: miR-96-5p; 150: miR-150; 183: miR-183-5p; 185: miR-185-5p; 194: miR-194-5p; 301: miR-320a; 371: miR-371a-5p; 3135: miR-3135b; 3652: miR-3652; 4534: miR-4534; 4698: miR-4698; 6088: miR-6088). Tubulin, control. (D) Migration assay using miR-96-5p, miR-150, miR-183-5p, miR-185-5p, miR-194-5p, miR-320a, miR-371a-5p, miR-3135b, miR-3652, miR-4534, miR-4698, miR-6088 and scrambled control (Scr) against My-La, HH, and HUT78 cells. RFUs (relative fluorescence units) were measured 16 hr after drug treatment. Student's t test was used for examining significance. Bars are means ± standard error of the mean (SEM) of three independent experiments. *0.01 ≤ P < 0.05, **0.001 ≤ P < 0.01, ***P < 0.001. Student's t-test was used to examine significance.