Journal: Journal of Thoracic Disease

Article Title: Bufalin inhibits epithelial-mesenchymal transition and increases radiosensitivity of non-small cell lung cancer via inhibition of the Src signaling

doi: 10.21037/jtd-22-1859

Figure Lengend Snippet: Bufalin inhibits EMT and abrogates radiation-induced EMT. (A) Western blot analysis for protein levels of E-cadherin, N-cadherin, vimentin, Slug, and Snail after treatment of A549 cells with increasing concentrations of bufalin. (B) Western blots to assess protein levels of E-cadherin, N-cadherin, vimentin, Slug, and Snail after treatment of A549 cells with DMSO, bufalin, radiation, and combined treatment (50 nM bufalin + radiation 4 Gy). (C) Cells were treated with radiation (4 Gy) or radiation plus bufalin; the changes of vimentin and E-cadherin in A549 cells were visualized by immunofluorescence microscopy. Scale bar, 20 µm. (D) Western blot analysis for protein levels of p-Y416 Src, Src (32G6), p-STAT3, and STAT3 after treatment of A549 cells with increasing concentrations of bufalin. (E) Representative immunofluorescence micrographs of A549 cells stained with antibodies against p-Y416 Src and p-STAT3. Scale bar, 20 µm. Ctrl, control; EMT, epithelial-mesenchymal transition; DMSO, dimethyl sulfoxide.

Article Snippet: The antibodies used in this study were purchased from Cell Signaling Technology (CST; Danvers, MA, USA), including anti-Src (#2123), anti-p-Y416-Src (#2102), anti-STAT3 (#12640), anti-p-STAT3 (#9145), anti-E-cadherin (#3195), anti-snail (#3879), anti-snail2 (#9585), anti-vimentin (#5741), anti-N-cadherin (#13116), anti-Vinculin (#4650), anti-GAPDH (#5174), and anti-β-actin (#3700).

Techniques: Western Blot, Immunofluorescence, Microscopy, Staining