Journal: Nature cancer

Article Title: LXRα activation and Raf inhibition trigger lethal lipotoxicity in liver cancer.

doi: 10.1038/s43018-020-00168-3

Figure Lengend Snippet: Fig. 2 | Sorafenib-mediated induction of LXRα signaling contributes to its therapeutic efficacy. a, Analysis of LXR activation in human HCCs upon sorafenib treatment. b, Results of IPA from mRNA-seq data of six human HCCs treated for 19–41 d with sorafenib (values represent z scores, canonical pathway analysis, comparison to gene expression at the start of treatment). c, mRNA expression analysis of Abcg5, Srebf1 and Scd1 in MycOE; NrasG12V HCCs in vivo upon treatment with sorafenib ± the reverse LXR agonist SR9238 (3 d, values represent the mean ± s.d., n = 6 mice per group, statistical significance was calculated by two-tailed Student’s t test; the analysis was repeated twice with similar results). d–f, Treatment of MycOE; NrasG12V HCCs with sorafenib or SR9238 alone, a combination thereof or the corresponding carrier. Representative pictures of livers following 5 weeks of treatment (e; n = 3 mice per group) and survival of mice under treatment (f; Kaplan–Meier curve, statistical significance was calculated by log-rank test, n = 6 mice per group). Scale bar, 1 cm. g–i, CRISPR-mediated editing of Nr1h3 in MycOE; NrasG12V HCCs and treatment of MycOE; NrasG12V + gNr1h3 or MycOE; NrasG12V + gNC HCCs with carrier or sorafenib. Representative pictures of livers following 5 weeks of treatment (h; n = 3 mice per group) and survival of mice under treatment (i; Kaplan–Meier curve, statistical significance was calculated by log-rank test, n = 6 mice per group). Scale bar, 1 cm. Numerical source data are provided.

Article Snippet: Ten percent of the supernatant was taken for input, and the rest was used for immunoprecipitation with 10 μl of Protein G beads (gel-agg-5, Invivogen) and antibodies to Raf-1 (Bethyl Laboratories, A301-519A; 1 μg), B-Raf (Sigma-Aldrich, HPA001328; 1 μg), SCD1 (Cell Signaling, 2794 (C12H5); 1 μg), FLAG (Sigma-Aldrich, F3165; 2.5 μg) or IgG control (Cell Signaling, 3900 (DA1E); 1 μg).

Techniques: Drug discovery, Activation Assay, Comparison, Gene Expression, Expressing, In Vivo, Two Tailed Test, CRISPR