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. Author manuscript; available in PMC: 2020 Mar 1.
Published in final edited form as: Cell Signal. 2018 Dec 19;55:26–39. doi: 10.1016/j.cellsig.2018.12.008

Fig. 1.

Fig. 1.

Cd induces increase in autophagosomes with a concomitant elevation of LC3-II and p62 in neuronal cells. PC12 cells and primary neurons, or PC12 cells and primary neurons infected with Ad-GFP-LC3 were treated with Cd (2.5, 5, 10 and/or 20 μM) for 2, 4, 6, 8, 10, 12 h and/or 24 h. (A and B) The cells were labeled using a specific autophagolysosome marker MDC staining and then the fluorescence intensity (in green) for MDC-labeled vacuoles was imaged (A) and quantified (B) as described in Materials and Methods. Scale bar: 20 μm. (C and D) Overlay of large LC3 puncta (in green) was shown (C) and the ratio of cells with large LC3 punctate structures was counted and calculated (D). Scale bar: 2 μm. (E and G) Total cell lysates were subjected to Western blotting using indicated antibodies. The blots were probed for β-tubulin as a loading control. Similar results were observed in at least three independent experiments. (F and H) The blots for LC3-II and p62 were semi-quantified. For (B), (D) (F) and (H), all data were expressed as means ± SE (n = 3–5). *P < 0.05, difference with control group.