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. 2021 May 1;12(5):437. doi: 10.1038/s41419-021-03708-6

Fig. 4. Lnc408 regulates the expression of target gene CBY1.

Fig. 4

A Schematic annotation of lnc408 genomic locus on chromosome 22. Black rectangles represent exons. B Intracellular localization of lnc408 was visualized in BC cells by RNA-FISH assays. Representative images of lnc408 in BT549 cells are shown. U6 served as a nucleus control and 18S as a cytoplasm control; cell nuclei were counterstained with DAPI (scale bar, 10 μm). C Full length of lnc408 transcripts (sense), antisense transcripts and control probe were labeled with biotin, then RNA-pull down was performed using nuclear extracts of mammospheres derived from BT549 cells. The interesting bands were followed by mass spectrometry. The arrow indicates the target of SP3. D The lnc408-binding protein SP3 in RNA-pull down precipitates (as carried out as in C, GAPDH as a negative control) was further confirmed by western blotting. E The interaction between lnc408 and SP3 was verified by an RNA immunoprecipitation (RIP) assay (**P < 0.01). Data are shown as means ± SD. F, G The mRNA or protein expression of CBY1 was determined by qRT-PCR (F) and western blotting (G) in lnc408-silenced BCSCs, respectively (*P < 0.05, **P < 0.01). H, I The mRNA and protein expressions of CBY1 were determined by qRT-PCR (H) and western blotting (I) in SP3-knocked down BCSCs, respectively (**P < 0.01).