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Fig. 6 | Molecular Cancer

Fig. 6

From: BAP1 dependent expression of long non-coding RNA NEAT-1 contributes to sensitivity to gemcitabine in cholangiocarcinoma

Fig. 6

BAP1 and NEAT-1 in CCA. a: Density plot of the distribution of BAP1 expression across 35 iCCA samples from The Cancer Genome Atlas (TCGA) database, along with b: expression levels of NEAT-1 RNA in patients with low versus high BAP1 expression. c: BAP1 and NEAT-1 mRNA expression by qRT-PCR in human CCA cell lines. d: Migration and invasion assays were performed in H69 cells as described in the Methods section. Illustrative bright field microscopic images are shown (left), and number of migrating (middle) or invasive (right) cells/field. e. The percentage change in viable cell number assessed using Cell Titer GloR 2.0 assay at the indicated times following knockdown of NEAT-1. f. Anchorage-independent growth of NEAT-1-transfected cells was assessed using a soft agar assay, with fluorometric assessment of colony formation after 7 days. Bars represent the mean + SEM of 6 separate studies. *, P < 0.05. g, h: BAP1 and NEAT-1 were exogenously modulated in KMBC cells to determine sensitivity to gemcitabine. KMBC cells were plated in a 6-well culture plate and after 12 h, medium was replaced with serum-free medium and cells were transfected with (G) NEAT-1 or non-targeting control siRNA for 6 h, or (H) with BAP1 or control plasmid for 24 h. Cells were then harvested, re-seeded at equal density in a 384-well plate and incubated with 25nM gemcitabine. Cytotoxicity was determined using Cell Titer GloR 2.0 assay after 72 h. Bars represent the mean + SEM of 4 separate determinants. *, P < 0.05

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