Figure 3.
Migfilin promotes autophagic flux. (A) KP4 cells were stably transfected with mRFP–GFP–LC3 report system. Representative images of mRFP-LC3 puncta and GFP-LC3 puncta in control (siCtrl/Vector), migfilin knockdown (siMig #1 or siMig #2), or migfilin overexpressed (Migfilin) KP4 cells under control condition. Scale bar, 10 µm (Magnify, 2 µm). Quantification of the percentage of RFP+GFP+ puncta to total puncta was shown in the right panel. Data represent mean ± SE, ***P < 0.001, one-way ANOVA. n = 3 independent experiments. At least 23 cells in each experiment were analyzed. (B) KP4 cells were stably transfected with mRFP-GFP-LC3 report system. Representative images of mRFP-LC3 puncta and GFP-LC3 puncta in control (siCtrl/Vector), migfilin knockdown (siMig #1 or siMig #2) or migfilin overexpressed (Migfilin) KP4 cells under starvation condition. Scale bar, 10 µm (Magnify, 1 µm). Quantification of the percentage of RFP+GFP+ puncta to total puncta was shown in the right panel. Data represent mean ± SE, **P < 0.01, ***P < 0.001, one-way ANOVA. n = 3 independent experiments. At least 29 cells in each experiment were analyzed. (C and D) SW1990 cells were stably transfected with mRFP–GFP–LC3 report system. Representative images of mRFP-LC3 puncta and GFP-LC3 puncta in control (siCtrl) or migfilin knockdown (siMig #1) SW1990 cells under both control (C) and starvation (D) condition. Scale bar, 10 µm (Magnify, 2 µm). Quantification of the percentage of RFP+GFP+ puncta to total puncta was shown in the right panel. Data represent mean ± SE, **P < 0.01, ***P < 0.001, unpaired two-tailed t test. n = 3 independent experiments. At least 35 cells in each experiment were analyzed.

Migfilin promotes autophagic flux. (A) KP4 cells were stably transfected with mRFP–GFP–LC3 report system. Representative images of mRFP-LC3 puncta and GFP-LC3 puncta in control (siCtrl/Vector), migfilin knockdown (siMig #1 or siMig #2), or migfilin overexpressed (Migfilin) KP4 cells under control condition. Scale bar, 10 µm (Magnify, 2 µm). Quantification of the percentage of RFP+GFP+ puncta to total puncta was shown in the right panel. Data represent mean ± SE, ***P < 0.001, one-way ANOVA. n = 3 independent experiments. At least 23 cells in each experiment were analyzed. (B) KP4 cells were stably transfected with mRFP-GFP-LC3 report system. Representative images of mRFP-LC3 puncta and GFP-LC3 puncta in control (siCtrl/Vector), migfilin knockdown (siMig #1 or siMig #2) or migfilin overexpressed (Migfilin) KP4 cells under starvation condition. Scale bar, 10 µm (Magnify, 1 µm). Quantification of the percentage of RFP+GFP+ puncta to total puncta was shown in the right panel. Data represent mean ± SE, **P < 0.01, ***P < 0.001, one-way ANOVA. n = 3 independent experiments. At least 29 cells in each experiment were analyzed. (C and D) SW1990 cells were stably transfected with mRFP–GFP–LC3 report system. Representative images of mRFP-LC3 puncta and GFP-LC3 puncta in control (siCtrl) or migfilin knockdown (siMig #1) SW1990 cells under both control (C) and starvation (D) condition. Scale bar, 10 µm (Magnify, 2 µm). Quantification of the percentage of RFP+GFP+ puncta to total puncta was shown in the right panel. Data represent mean ± SE, **P < 0.01, ***P < 0.001, unpaired two-tailed t test. n = 3 independent experiments. At least 35 cells in each experiment were analyzed.

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