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. 2011 Feb 1;7(2):176–187. doi: 10.4161/auto.7.2.14074

Figure 1A–G.

Figure 1A–G

Caffeine increases autophagic flux in various cell lines. (A) structural formula of caffeine. (B and C) SH-SY5Y cells treated with various concentrations of caffeine for 24 or 48 hours were analyzed by immunoblotting (B) with antibodies against LC3 and actin. Densitometry analysis of LC3-II levels relative to actin (C) was performed using three independent experiments. (D and E) SH-SY5Y cells treated with 25 mM caffeine for 3–24 hours were analyzed by immunoblotting (D) with antibodies against LC3 and actin. Densitometry analysis of LC3-II levels relative to actin (E) was performed using three independent experiments. (F) Electron microscopic examination of SH-SY5Y cells treated with various concentrations of caffeine for 24 or 48 hours. Autophagic vacuoles accumulating in the cytoplasm are shown by arrows. (G) Morphometric analysis of autophagic vacuoles was performed with 30 different areas of the cytoplasm of control and caffeine-treated cells.