Skip to main content
. 2021 Apr 13;11:610885. doi: 10.3389/fonc.2021.610885

Figure 6.

Figure 6

Cultivation of A2780 (A, C, D) and SKOV3 (B, E) OC cells in lipid-free media inhibits growth (A, B), down-regulates expression of lipid metabolic proteins (C) and conditions the cells for restorative import of exogenously supplied FAs or LDLs (D, E). Cells were cultured for 0 – 4 days (A, left panel) or for 72 hours (A, right panel; B both panels) in media containing 5% FCS with or without lipids. Analysis of cell growth was either by formazan dye assay (A both panels; B left panel) or direct microscopic counting in a hemocytometer (B right panel). Western blots (C) were (co-)probed for lipid metabolic proteins, stripped if necessary, and re-probed for actin as loading control. The band intensities obtained after chemoluminescence detection of autoradiographs were determined using ImageJ software and the ratios between the proteins of interest and the corresponding actin on the same membrane were calculated. Horizontal lines separate the different membranes. These ratios were normalized to cultures grown for 48 hours in media supplemented with 5% FCS + lipids (arbitrarily set to 100%) and are given under each autoradiograph in percent. Autoradiographs of one of two experiments are shown. For further technical details of cell proliferation experiments, Western blotting, and uptake assays for FAs and LDLs see Materials and methods. Means ± SD, n = 3. Student’s t-test (A, B) or ANOVA followed by Scheffe test (D, E), p < 0.05 (*), p < 0.01 (**) and p < 0.001 (***).