Skip to main content
. 2020 Jun 23;31(12):107806. doi: 10.1016/j.celrep.2020.107806

Figure 4.

Figure 4

Modulation of SREBP Alters FADS2 Transcription and Sapienate Metabolism

(A and D) Relative FADS2 (A) and SCD1 (D) mRNA expression relative to actin beta (ACTB) was determined in U87 cells stably expressing ER-mSREBP-1 or ER-mSREBP-2. Cells were treated with 100 nM 4-OHT or solvent for 24 h in medium containing 10% FBS (n = 3). Bar graphs are presented as mean ± SD from biological independent samples. Statistical testing was performed by a two-sided unpaired Student’s t test.

(B and E) Sapienate to palmitate ratio (B) and palmitoleate to palmitate ratio (E) were determined in U87 cells stably expressing ER-mSREBP-1 or ER-mSREBP-2. Cells were treated with 100 nM 4-OHT or solvent for 24 h in medium containing 10% FBS (n = 3). Bar graphs are presented as mean ± SD from biological independent samples. Statistical testing was performed by a two-sided unpaired Student’s t test.

(C and F) Relative sapienate (C) and palmitoleate (F) levels in U87 cells stably expressing ER-mSREBP-1 or ER-mSREBP-2 (n = 3). Bar graphs are presented as mean ± SD from biological independent samples. Statistical testing was performed by a two-sided unpaired Student’s t test.

(G) Relative FADS2 mRNA expression upon treatment with DMSO (n = 10) or fatostatin (10 μM, 72 h, WT n = 11, Tsc2−/− n = 10, HUH7 n = 9) in WT and Tsc2−/− MEFs or HUH7 cancer cells. Bar graphs are presented as mean ± SD from biological independent samples. Statistical testing was performed by a two-sided unpaired Student’s t test.

(H) Sapienate to palmitate ratio upon treatment with DMSO (n = 12) or fatostatin (10 μM, 72 h, MEFs n = 12, HUH7 n = 11) in WT and Tsc2−/− MEFs or HUH7 cancer cells. Bar graphs are presented as mean ± SD from biological independent samples. Statistical testing was performed by a two-sided unpaired Student’s t test.

(I) Relative sapienate level upon treatment with DMSO (n = 12) or fatostatin (10 μM, 72 h, MEFs n = 12, HUH7 n = 11) in WT and Tsc2−/− MEFs or HUH7 cancer cells. Bar graphs are presented as mean ± SD from biological independent samples. Statistical testing was performed by a two-sided unpaired Student’s t test.

(J and M) Relative FADS2 (J) and SCD1 (M) protein expression upon treatment with DMSO or fatostatin (10 μM, 72 h, n = 3) in HUH7 cancer cells. Bar graphs are presented as mean ± SD from biological independent samples. Statistical testing was performed by a two-sided unpaired Student’s t test.

(K) Relative SCD1 mRNA expression in WT and Tsc2−/− MEFs or HUH7 cancer cells upon treatment with DMSO (MEFs n = 6, HUH7 n = 11) or fatostatin (10 μM, 72 h, MEFs n = 7, HUH7 n = 9). Bar graphs are presented as mean ± SD from biological independent samples. Statistical testing was performed by a two-sided unpaired Student’s t test.

(L) Palmitoleate to palmitate ratio in WT and Tsc2−/− MEFs or in HUH7 cancer cells upon treatment with DMSO (n = 12) or fatostatin (10 μM, 72 h, MEFs n = 12, HUH7 n = 11). Bar graphs are presented as mean ± SD from biological independent samples. Statistical testing was performed by a two-sided unpaired Student’s t test.