Fig. 3: Autophagy is required in the progress of ferritinophagy induced by 25-HC. | Cell Death Discovery

Fig. 3: Autophagy is required in the progress of ferritinophagy induced by 25-HC.

From: Oxysterol 25-hydroxycholesterol activation of ferritinophagy inhibits the development of squamous intraepithelial lesion of cervix in HPV-positive patients

Fig. 3

A Bright field images of the H8 cells and SiHa cells treated with or without 25-HC (10 μM). Scale bar, 50 μm. B The H8 cells and SiHa cells were treated with or without 25-HC for 24 h. Then they were fixed and immune-stained with anti-LC3 (green) and anti-LAMP1 (red). Scale bar, 50 μm (inserts, 10 μm). C The H8 cells and SiHa cells were treated with or without 25-HC for 24 h. Then they were fixed and immune-stained with anti-LC3 (green) and anti-FTH1 (red). Scale bar, 50 μm (inserts, 10 μm). D TEM images of ultrastructure in the H8 cells and SiHa cells treated with or without 25-HC. MT, mitochondria; AL, autolysosome; LY, lysosome; Scale bar, 5 μm (inserts, 1 μm). E Lysates of the H8 cells and SiHa cells treated with or without 25-HC for 24 h were immunoblotted for NCOA4, FTH1 and ACTB (n = 3). The data are normalized to the ACTB control. F The H8 cells and SiHa cells were treated with wortmannin or chloroquine for 1 h and continued cultured adding 25-HC for 24 h. Then they were fixed and stained with FerroOrange (red) to identify Fe (II). Scale bar, 50 μm. G Lysates of the H8 cells and SiHa cells treated with wortmannin or chloroquine were immunoblotted for LC3B, NCOA4, FTH1 and ACTB (n = 3). The data are normalized to the ACTB control. All data are from three independent experiments. The data are presented as the mean ± SD values (n ≥ 3). ***P < 0.001.

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