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. Author manuscript; available in PMC: 2012 May 1.
Published in final edited form as: Biochem Pharmacol. 2011 Mar 1;81(9):1152–1163. doi: 10.1016/j.bcp.2011.02.015

Figure 1. EGCG was internalized into cytoplasmic vesicles containing LC3, LAMP2, and HMGB1.

Figure 1

A). EGCG was co-localized with LC3-containining vesicles. GFP-LC3-transfected RAW 264.7 cells were treated with biotin-labeled EGCG for 6 h, subsequently stained with streptavidin-conjugated Alexa-594 to visualize intracellular EGCG, and stained with DAPI to visualize the nuclei (blue). B). EGCG was co-localized with LAMP2-positive vesicles. RAW 264.7 cells were treated with biotin-labeled EGCG for 16 h, and stained with LAMP2-specific primary antibody and Alexa-488-conjugated secondary antibody. C). EGCG co-localized with HMGB1-containing cytoplasmic vesicles. RAW 264.7 cells were stimulated with LPS in the presence of biotin-EGCG for 16 h, and stained HMGB1-specific antibodies, and Alexa-488-conjugated donkey anti-rabbit antibody. D). Cytoplasmic HMGB1 “puncta” were partly co-localized with LC3-containining vesicles. GFP-LC3-transfected RAW 264.7 cells were stimulated with LPS and EGCG for 6 h, and sequentially incubated with HMGB1-specific and Alexa-594-conjugated donkey anti-rabbit antibodies. Note that cytoplasmic HMGB1 “puncta” (Red) partly co-localized with LC3-containing cytoplasmic vesicles (Green).