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. 2018 Sep 4;24(10):2596–2605.e5. doi: 10.1016/j.celrep.2018.08.015

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© 2018 The Author(s)

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

PMC Copyright notice

DGAT Loss Modifies Lipid Homeostasis, Elevates Ceramide, Acyl-ceramide, and Acyl-carnitine Levels, and Activates NF-kB Target Gene Expression

(A) Effect of SCD and DGAT inhibition on ceramide levels in serum-deprived A498 cells in vitro.

(B) Effect of DGAT loss on ceramides in vivo (i.e., A498 xenografts).

(C) Effect of DGAT loss on acyl-ceramides in vivo (i.e., A498 xenografts).

(D) Effect of hypoxia on the FA composition of acyl-carnitines (CARs) on serum-deprived A498 cells in vitro.

(E) Effect of DGAT loss on the FA composition of acyl-carnitines (CARs) on serum-deprived A498 cells in vitro.

(F) Effect of DGAT loss on the FA composition of acyl-CARs in A498 xenograft tumors.

(G) Gene set enrichment analysis (GSEA) on RNA as assessed by microarray comparisons performed on A498 DGAT shRNA xenograft tumors after 5 days of control or doxycycline chow. Normalized enrichment score (NES) allow comparison of enrichment between different gene sets.

(H) Effect of DGAT shRNA, NF-κB inhibition, and proteasome inhibition on NF-κB luciferase reporter activity.

(I) Effect of serum deprivation on NF-κB luciferase reporter activity.

(J) Schematic of the consequences of DGAT inhibition preceding a period of unsaturated lipid deprivation. These conditions lead to increased acyl-CARs and ceramides as well as increased incorporation of saturated FAs into the PL pool.

(K) Published data comparing the TG composition of ccRCC and normal tissue (Saito et al., 2016) were reanalyzed to investigate shifts in TG saturation. TG species were binned according to the number of fully saturated FA chains present, and the abundance of each category was aggregated and is displayed as a ratio of the abundance in normal tissue.

For (A), (D), (E), data are means of 5 and for (H) and (I) of 3 replicate wells, and results were confirmed in independent experiments. For (B), (C), (F), and (G), data are means of tumors from 4 tumors. Error bars represent SD. Statistical significance by t test or ANOVA, as appropriate. ^∗p < 0.05, ^∗∗p < 0.05, and ^∗∗∗p < 0.005. See also Figure S4.