Fig. 2

Effect of HOTAIR overexpression in c-Met downstream signaling. Analysis of c-Met and downstream signaling in MOCK and HOTAIR OE cells. Immunoblot analysis of (a) c-Met protein expression, c-Met activation phosphorylation (Tyr-1234/Tyr-1235); (b) Akt1 protein expression, Akt1 activation phosphorylation (Ser-473); (c) Erk1/2 protein expression, Erk1/2 activation phosphorylation (Thr-202/Tyr-204); (d) STAT3 protein expression and activation phosphorylation (Tyr-705). Column graph of corrected total cell fluorescence of c-Met immunofluorescent staining (e). Immunofluorescence imaging of (f) total c-Met protein (red) and its activation (green). RT- qPCR analysis of (g) CAV1 mRNA expression, immunoblotting of (h) Caveolin-1 protein and its activation phosphorylation (Tyr-14) and column graph of densitometric analysis of Caveolin-1 activation and total protein expressions. Immunoblotting of (i) Src kinase expression and its activation phosphorylation (Tyr-416). Immunofluorescence imaging of (j) total Caveolin-1 protein (red) and its activation (green). DAPI (blue) was used as a nuclear marker in immunofluorescent images. Densitometric analysis of band intensities in immunoblots were analyzed with ImageJ and normalized with internal control protein band intensities. 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