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. 2023 Aug 23;42(40):2956–2970. doi: 10.1038/s41388-023-02814-3

Fig. 7. BLACAT3 binds and induces YBX3 to shuttle into the nucleus.

Fig. 7

A RNA pull-down and silver staining showed the molecular weight of BLACAT3 binding proteins. B WB assay using the protein sample pulled by BLACAT3 proved that YBX3 is the binding protein of BLACAT3. C, D RNA Immunoprecipitation (RIP) and qRT-PCR assay confirmed that BLACAT3 can bind with YBX3 protein. E, H Western blot demonstrated that BLACAT3 knockdown had no effect on YBX3 protein levels in T24 and 5637 cells. F, I The effect of BLACAT3 knockdown on the subcellular distribution of YBX3 in T24 cells was detected by WB assay after separation of nuclear and cytoplasmic proteins. G, J The effect of BLACAT3 overexpression on YBX3 subcellular distribution in 5637 cells was detected by WB assay after separation of nuclear and cytoplasmic proteins. K NSG mice were subcutaneously injected with T24 cells with BLACAT3 stable knockdown, and the mice were sacrificed on the 28th day. The subcutaneous tumor was isolated, fixed and embedded, and then H&E staining and anti-YBX3 immunofluorescent (IF) staining assessed the effect of BLACAT3 knockdown on the subcellular distribution of YBX3. Scale bars: 1 mm (black and white lines), 20 μm (orange lines). ns no significance. Statistical significance was assessed using two-tailed Student’s t test between two groups, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.