Fig. 3
Analysis of HOTAIR-c-Met interaction in MET overexpressed, activated and inhibited conditions. Immunoblotting of (a) c-Met activation phosphorylation (Tyr-1234/Tyr-1235), and total c-Met protein expression. Confocal microscopy imaging of (b) c-Met protein and RT-qPCR analysis of c-Met mRNA expression in MET OE SNU-398 clones. (c) Confocal imaging of FISH labeling (white dots) and RT-qPCR analysis of HOTAIR mRNA expression in MET OE clones. (d) Confocal imaging of Caveolin-1 protein and qPCR analysis of CAV1 mRNA expression in MET OE cells. (e) Immunoblot of c-Met activation and qPCR analysis of HOTAIR expression in ligand-dependent activation (HGF 10 ng/ml) of c-Met in HuH-7, HEP-3B, SNU-449, MAHLAVU and SK-HEP-1 cells. RT-qPCR analysis of HOTAIR expression (f), in response to heparin (100 μg/mL) induced c-Met activation in SK-HEP-1 cells. RT-qPCR analysis of MET and HOTAIR expressions in sorafenib resistance (MAHLAVU cell line) (g) and high-glucose (25 mM, HuH-7 cell line) (h) and fluidic shear stress (0.5 dyn/cm2, HuH-7 cell line) (i) induced c-Met activations. (j) Immunoblot of c-Met activation and qPCR analysis of HOTAIR expression in tyrosine kinase activity inhibition of c-Met by SU11274 (2500 nM) HuH-7, SNU-449, MAHLAVU and SK-HEP-1 cells. All graphs of experiments are presented as the mean ± SEM of at least 3 independent experiments. * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001