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. 2019 May 3;20(9):2192. doi: 10.3390/ijms20092192

Figure 3.

Figure 3

Effect of copper and zinc acetate treatment on autophagy. (A) Huh7 cells were treated with copper at the concentration of 300 μM for 12 h, and zinc acetate (200 μM) was added 2 h before copper sulfate treatment. Densitometry analysis of indicated proteins is shown on the bottom. (B) OUMS-29 cells were transfected with monomeric red fluorescent protein (mRFP)–green fluorescent protein (GFP) tandem fluorescent-tagged light chain 3 (LC3) plasmids. They were treated with copper at concentrations of 100 or 300 μM for 12 h, and zinc acetate (200 μM) was added 2 h before copper sulfate treatment. Colocalizations of mRFP and GFP signals were measured by counting an overall total of 30 to 40 cells over the course of three independent experiments. Colocalization was shown as the percentage of merged signals in the total number of mRFP signals. Data are expressed as means ± SEM. (C) Huh7 cells were treated with zinc acetate at concentrations of 100 or 200 μM for 2 h. Densitometry analysis of indicated proteins is shown on the bottom. (D) Huh7 cells were treated with 4-phenylbutyrate (PBA) at the concentration of 5 mM for 2 h. Densitometry analysis of indicated proteins is shown on the right. (E) Huh7 cells were treated with copper at the concentration of 300 μM for 12 h, and PBA (5 mM) was added 2 h before copper sulfate treatment. Densitometry analysis of indicated proteins is shown on the right. (F) OUMS-29 cells were transfected with mRFP–GFP tandem fluorescent-tagged LC3 plasmids. They were treated with copper at the concentration of 300 μM for 12 h, and PBA (5 mM) was added 2 h before copper sulfate treatment. Colocalizations of mRFP and GFP signals were measured by counting an overall total of 30 to 40 cells over the course of three independent experiments. Colocalization was shown as the percentage of merged signals in the total number of mRFP signals. Data are expressed as means ± SEM. (G) Huh7 cells were treated with zinc acetate at the concentration of 200 μM for 2 h. Densitometry analysis of indicated proteins is shown on the right. (H) Huh7 cells were treated with copper at the concentration of 300 μM and tunicamycin at the concentration of 0.5 μM for 12 h, and zinc acetate (200 μM) was added 2 h before copper sulfate treatment. Densitometry analysis of indicated proteins is shown on the right.