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

Figure 2.

Figure 2

mTOR Can Regulate Fads2 Expression and Sapienate Metabolism

(A) Representative western blot of mTORC1 activity in wild-type (WT) and Tsc2−/− MEFs. mTORC1 activity was analyzed by immunoblotting with the indicated antibodies.

(B) Relative Fads2 mRNA expression in WT (n = 11) and Tsc2−/− (n = 12) MEFs based on qRT-PCR. 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) Relative sapienate to palmitate ratio in WT and Tsc2−/− MEFs based on mass spectrometry analysis. n = 18. Bar graphs are presented as mean ± SD from biological independent samples. Statistical testing was performed by a two-sided unpaired Student’s t test.

(D and E) Relative Fads2 mRNA expression in WT and Tsc2−/− MEFs upon treatment with DMSO (D: WT n = 11, Tsc2−/− n = 10; E: WT n = 8, Tsc2−/− n = 7), rapamycin (D) (20 nM, 72 h, WT n = 9, Tsc2−/− n = 10), or Torin1 (E) (40 nM, 72 h, WT n = 9, Tsc2−/− n = 7). Bar graphs are presented as mean ± SD from biological independent samples. Statistical testing was performed by a two-sided unpaired Student’s t test.

(F and G) Sapienate to palmitate ratio in WT and Tsc2−/− MEFs upon treatment with DMSO (F: WT N = 11, Tsc2−/− n = 12; G: WT n = 11, Tsc2−/− n = 10), rapamycin (F) (20 nM, 72 h, WT n = 10, Tsc2−/− n = 12), or Torin1 (G) (40 nM, 72 h, WT n = 11, Tsc2−/− 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.

(H and I) Relative sapienate level in WT and Tsc2−/− MEFs upon treatment with DMSO (H: n = 12; I: n = 11), rapamycin (H) (20 nM, 72 h, n = 12), or Torin1 (I) (40 nM, 72 h, 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.