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Figure 4 | Molecular Cancer

Figure 4

From: Phospholipid Scramblase 1, an interferon-regulated gene located at 3q23, is regulated by SnoN/SkiL in ovarian cancer cells

Figure 4

PLSCR1 modulates As 2 O 3 -induced cell death response in HEY ovarian carcinoma cells. (A) HEY cells were treated with 25 μM As2O3 (15 min - 24 h) followed by harvesting of cell lysates and analyses via western blotting for the indicated proteins. Results are representative of triplicate experiments. (B) Densitometric analysis of PLSCR1 protein levels of data presented in (A). (C) HEY cells were treated with 25 μM As2O3 (6 or 18 h) in the absence or presence of MG132 (5 μM) followed by harvesting of cell lysates and analyses via western blotting for the indicated proteins. Results are representative of duplicate experiments. (D) HEY cells were treated with 25 μM As2O3 (6 or 18 h). RNA was isolated and analyzed for PLSCR1 mRNA levels by real-time PCR. Results are representative of duplicate experiments. (E) Cell lysates were collected from HEY cells treated with non-targeting or PLSCR1 siRNA in the absence or presence of 25 μM As2O3 (18 h). Western blotting was performed for the proteins indicated. Results are representative of triplicate experiments. (F) Cells were transfected with pEGFP-LC3. Twenty-four hours post transfection, cells were treated with non-targeting and PLSCR1 siRNA followed by treatment with 10 μM As2O3. Immunofluorescence was performed (left panel) and the data was quantitated (bar graphs, right panel) by counting the number of cells positive for EGFP-LC3 punctae. Results are representative of duplicate experiments. (G) A schematic showing regulation of PLSCR1 mRNA by SnoN and As2O3 (and its effect on the cell death response) in HEY cells. In addition to the downstream effect of overexpressed PLSCR1 on PAI-1, a TGFβ target gene, in T80 cells.

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