Journal: Biochemical and biophysical research communications

Article Title: TMEM2 expression is downregulated as bladder cancer invades the muscle layer.

doi: 10.1016/j.bbrc.2022.04.118

Figure Lengend Snippet: Fig. 3. TMEM2 expression decreases in EMT-induced cells. A) Non-invasive BCa RT4 cells were cultured 4 days with (panels b, d) or without (panels a, c) StemXVivo EMT Inducing Media Supplement (x2). On day 4, cells were analyzed for epithelial phenotypes both by bright field imaging (panels a, b; Scale Bars: 100 mm) and staining with anti-human E-cadherin (green) (panels c, d; Scale Bars: 50 mm). B) Fluorescence intensity calculated for E-cadherin, with (þ) or without () EMT Inducing Media Supplement. C) TMEM2 fluorescence intensity quantified in RT4 cells with (þ) or without () EMT Inducing Media Supplement. *****; significant difference between groups P < 0.0001. D) TMEM2 immunofluorescence (green) in RT4 cells with (þ) or without () EMT Inducing Media Supplement. Nuclei are stained with DAPI (blue). Without supplementation, TMEM2 is expressed at the cell membrane (D- a, arrow); with supplementation, TMEM2 is detected in a pattern of dots around the nucleus (D-b, arrow). Scale Bars: 10 mm. . (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: To induce the EMT, RT4 cells were grown 4 days in RPMI1640 with 10% FBS supplemented with EMT induction supplement (x2) (StemXVivo EMT Inducing Media Supplement, R&D systems, Minneapolis, MN, USA), which contains anti-human E-cadherin, anti-human sFRP-1 and anti-human Dkk-2 antibodies, as well as recombinant human Wnt-5a and human TGF-b proteins [23].

Techniques: Expressing, Cell Culture, Imaging, Staining, Fluorescence, Membrane