Fig. 4: LINC00880 interacts with CDK1 to enhance its kinase activity. | Cell Death & Disease

Fig. 4: LINC00880 interacts with CDK1 to enhance its kinase activity.

From: The super-enhancer-driven lncRNA LINC00880 acts as a scaffold between CDK1 and PRDX1 to sustain the malignance of lung adenocarcinoma

Fig. 4

A Identification of LINC00880-interacting proteins. LINC00880 and the antisense of LINC00880 were synthesized and biotinylated in vitro followed by the RNA pull-down assay. The silver staining reveals the specific bands that were subjected to MS analysis (indicated with arrow). B LINC00880 RNA pulldown-MS candidates sorted for log-rank P-value in TCGA-LUAD patient survival analysis. C, D The interaction between LINC00880 and CDK1 was determined by a RIP assay using a CDK1-specific antibody, immunoglobulin G (IgG) was used as the negative control. C: PC9, D: H358. E, F By the RNA pull-down assay, CDK1 is pulled down with LINC00880, but not with the antisense of LINC00880. E: PC9, F: H358. G LINC00880 is predicted to have four stem-loop structures by using Rfold. H Immunoblot detection of the CDK1 protein in PC9 cells as retrieved by in vitro transcribed biotinylated RNAs of different constructs of LINC00880 or its antisense sequence (as negative control). I, J Activity of CDK1 in cell lysates was measured using a Histone H1 kinase substrate. I: PC9, J: H358. K, L Western blotting detection of cyclin B and phosphorylated CDK1 (T14/Y15/T161) expression in PC9 cells transfected with pcDNA3.1, LINC01977, si-NC, si-LINC00880. K: PC9, L: H358. M, N Co-immunoprecipitation (co-IP) of CDK1 with CAKs in PC9 cells transfected with LINC00880. M: PC9, N: H358. The data are shown as the mean ± S.D. of at least three replicates (*P < 0.05, **P < 0.01, ***P < 0.001).

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