Fig. 5

lncBCL2L11 interacts with THOC6 and regulates THOC5 stability. A FISH assay was performed to exam the location of lncBCL2L11 in GBC tissues. Scale bar, 100 μm. B RNA pull down assay by lncBCL2L11 and its antisense RNA followed by silver staining. Differential protein band was excised for mass spectrometry analysis. Western blot analysis revealed the specific binding between lncBCL2L11 and THOC6.ELAV1 used as negative control. C Immunoblotting of THOC6 in pull-down samples by biotin labeled anti-sense lncBCL2L11 (F1), full-length lncBCL2L11 (F2) and truncated lncBCL2L11 framents (F3:1-400nt; F4:1-200nt; F5:400-631nt). D RIP assay was performed using anti-THOC6 antibody in NOZ extracts followed by RT-PCR. E, F Protein levels of THOC2, THOC5, THOC6 after lncBCL2L11 overexpression and knockdown. G Interaction between THOC5 and THOC6 was detected by CO-IP assay. H, I CO-IP assay was performed to detected the interaction between THOC6 and THOC5 after endogenous or exogenous increase or decrease of lncBCL2L11. J Western blot of proteins from control and lncBCL2L11 overexpression NOZ cells after adding actinomycin to prevent protein synthesis