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Fig. 7 | Cancer Cell International

Fig. 7

From: AL355338 acts as an oncogenic lncRNA by interacting with protein ENO1 to regulate EGFR/AKT pathway in NSCLC

Fig. 7

AL355338 activated the EGFR/AKT pathway by targeting ENO1/EGFR interaction. A 293T cells were co-transfected with flag-ENO1 and HA-EGFR. Reciprocal immunoprecipitation was performed with anti-HA or anti-flag agarose (IgG as control). Input (whole-cell extracts) and immunoprecipitates were analyzed by SDS-PAGE and immunoblotted with indicated antibodies. B Lysates of A549 cells were immunoprecipitated with the antibody against ENO1 or EGFR. Input and immunoprecipitates were analyzed by SDS-PAGE and immunoblotted with indicated antibodies. C Subcellular co-localization of ENO1 and EGFR in A549 and PC9 cells. Cells were immunostained with anti-ENO1 (red) and anti-EGFR (green). The nucleus was marked with DAPI (blue). Scale bar, 10 μm. D In situ proximity ligation assay (PLA) demonstrated the interaction between ENO1 and EGFR in A549 and PC9 cells. Positive PLA signals demonstrated ENO1/EGFR complex which were shown as red clusters, and cell nucleus were counterstained with DAPI (blue). Scale bar, 10 μm. E, F The interaction between ENO1/EGFR was detected in AL355338-knockdown A549 cells and AL355338-overexpressing PC9 cells using co-IP assays. G Western blotting analysis of p-EGFR, EGFR, p-AKT, AKT in A549 and PC9 cells treated with sh-AL355338 alone or co-transfection with ENO1 overexpression plasmid. H Western blotting analysis of p-EGFR, EGFR, p-AKT, AKT in A549 and PC9 cells treated with AL355338 overexpression alone or co-transfection with sh-ENO1

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