Research Paper Volume 12, Issue 24 pp 25845—25864

The long noncoding RNA LINC01140/miR-140-5p/FGF9 axis modulates bladder cancer cell aggressiveness and macrophage M2 polarization

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Figure 6.

Dynamic effects of LINC01140 and miR-140-5p on bladder cancer cell aggressiveness and macrophage M2 polarization T24 cells were cotransfected with si-LINC01140 and anti-miR-140-5p and examined for (A) the cell viability by MTT assay; (B) protein levels of FGF9, ki-67, MMP-2, and MMP-9 by immunoblotting; (C) migration capacity by wound healing assay; and (D) invasive capacity by Transwell assay. (E, F) T24 cells were cotransfected with si-LINC01140 and anti-miR-140-5p and the culture medium (shown in the figures as conditioned medium (CM) si-NC + anti-NC, si-LINC01140 + anti-NC, si-NC + anti-miR-140-5p, and si-LINC01140 + anti-miR-140-5p) was collected for macrophage incubation. M0 macrophages were cultured in the above-described four kinds of CMs and polarized to M1 or M2, respectively, and examined for (E) the protein levels of CD206 and CD16 by immunoblotting. (F) The concentrations of IL-10, Arg1, iNOS, and TNF-α in the macrophage culture medium was determined by ELISA. *P<0.05, **P<0.01, compared to control group; #P<0.05, ##P<0.01, compared to si-NC + anti-miR-140-5p group. One way ANOVA test was used for statistical analysis.