Journal: Journal of Cellular and Molecular Medicine
Article Title: Blue LED causes autophagic cell death in human osteosarcoma by increasing ROS generation and dephosphorylating EGFR
doi: 10.1111/jcmm.16412
Figure Lengend Snippet: Blue LED irradiation prevents EGFR activation (phosphorylation) and EGFR/Beclin‐1 complex formation. U‐2 OS cells were subject to three types of red/green/blue LED irradiation for 180 J/cm 2 and 360 J/cm 2 . A, The expression of pEGFR was detected by immunostaining promptly. Representative images were captured with a confocal microscopy. (Bar: 50 μm). B, The protein levels of total EGFR and pEGFR after treatment of blue LED. C, Colocalization of EGFR (Green) and Beclin‐1 (Red) was detected by immunostaining. The yellow spots indicate colocalization for both. (Bar: 50 μm). D, Validation of Beclin‐1 and EGFR combination in U‐2 OS cells detected by co‐immunoprecipitation (Co‐IP) with anti‐EGFR or anti‐IgG. Data are expressed as the mean ± SEM. ** P < 0.01; *** P <0.001
Article Snippet: The following primary antibodies and concentrations were used: Beclin‐1 (1:1000, cat. no. 3738S, CST), LC3 (1:1000, cat. no. L7543, Sigma‐Aldrich, St. Louis, Missouri, United States), p62 (1:1000, cat. no. 5114S, CST), EGFR (1:500, cat. no. Bs‐10007R, BIOSS, Bioss Antibodies, Boston, Massachusetts, United States), pEGFR (Tyr1172, 1:500, cat. no. Bs‐20385R, BIOSS), NOX2 (1:1000, cat. no. ab129068, Abcam), NOX4 (1:1000, cat. no. 14347‐1‐AP, Proteintech), Catalase (CAT, 1:2000, cat. no. ab209211, Abcam) and β‐actin (1:1000, cat. no. 8457S, CST).
Techniques: Irradiation, Activation Assay, Expressing, Immunostaining, Confocal Microscopy, Immunoprecipitation, Co-Immunoprecipitation Assay
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