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Fig. 7 | Journal of Experimental & Clinical Cancer Research

Fig. 7

From: TFE3 fusions escape from controlling of mTOR signaling pathway and accumulate in the nucleus promoting genes expression in Xp11.2 translocation renal cell carcinomas

Fig. 7

TFE3 fusions regulate lysosome biogenesis genes. The mRNA expression of TFE3, SQSTM1, VPS8, VPS11 ATP5G, ATP5O, 4EBPs and GOLPH3 were detected by q-PCR. Western blot was used to detect the expression of TFE3, 4EBP3, S6, and p-S6 on protein level. Immunofluorescence analysis were applied to confirm the subcellular localization of TFE3 fusion proteins. (a, b) UOK109 and UOK120 cells were transfected with TFE3 knockdown shRNA (Y3619 and Y3620) or negative control shRNA (Y007) for 48 h. mRNA levels of TFE3 and other related genes were examined by q-PCR. (c, d) UOK109 and UOK120 cells were transfected with TFE3 knockdown virus (54,157, 54,158 and 54,159) or negative control virus (CON077) for 72 h. mRNA levels of TFE3 and other related genes were detected with q-PCR. (e) UOK109 cells were transfected with 54,157 (TFE3-KD) or CON077 (TFE3-NC). After 72 h incubation, TFE3 fusions (red) were stained with antibodies and measured with immunofluorescence. Nucleus were stained with Hoechst (blue) and EGFP showed expression intensity of the virus. Scale bar, 50 μm. (f) UOK120 cells were transfected with 54,157 (TFE3-KD) or CON077 (TFE3-NC). After 72 h incubation, TFE3 fusions (red) were stained with antibodies and measured with immunofluorescence. Nucleus were stained with Hoechst (blue) and EGFP (green) showed expression intensity of the virus. Scale bar, 50 μm. Data were presented as the mean ± SEM. *P < 0.05, **P < 0.01. The unpaired two-tailed Student’s t-test was used to calculate P value

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