acss2  (MedChemExpress)

Journal: Cell reports

Article Title: KRAS G12V mutation-selective requirement for ACSS2 in colorectal adenoma formation

doi: 10.1016/j.celrep.2025.115444

Figure Lengend Snippet: (A) Representative western blot for expression and activation of lipogenic enzymes in mouse colon epithelial cells cultured in 0.5% FBS and upon stimulation with EGF for 20 min. Results are representative of three similar experiments. (B) Quantification of western band intensities in (A) for ACSS2. One-way ANOVA; ns, not significant; * p < 0.05, ** p < 0.01. (C) Intracellular cholesterol ester levels in KRAS-mutant cells. Data show Cholesterol Ester-Glo luminescence averages from three biological replicates. One-way ANOVA; ns, not significant; * p < 0.05, **** p < 0.0001. (D) Experimental design for heavy isotope labeling of acetyl-CoA using [U- 13 C] glucose with unlabeled acetate (left) and [1,2- 13 C] acetate with unlabeled glucose (right). (E) m + 2 acetyl-CoA following 24-h labeling with 5 mM [U- 13 C] glucose with 1 mM unlabeled acetate (left) and 1 mM [1,2- 13 C] acetate with 5 mM unlabeled glucose; ** p < 0.01, **** p < 0.0001. (F) Representative western blot confirming ACSS2 KO using CRISPR. (G) Intracellular cholesterol ester levels in KRAS-mutant; ACSS2 KO cells. Data show Cholesterol Ester-Glo luminescence averages from three biological replicates. One-way ANOVA; ns, not significant; **** p < 0.0001. (H) Representative western blot of ACSS2 subcellular localization in KRAS-mutant cells. (I) Quantification of western band intensities in (H) for cytoplasmic ACSS2. One-way ANOVA; ** p < 0.01, **** p < 0.0001. (J) ACSS2-dependent acetylation of RAPTOR in KRAS G12V CRC cells. Proteins were immunoprecipitated with anti-acetylated lysine antibodies (Abs), and the immunoprecipitates (IPs) were probed with anti-RAPTOR Abs.

Article Snippet: ACSS2 inhibitor , Selleck , Cat# S8588.

Techniques: Western Blot, Expressing, Activation Assay, Cell Culture, Mutagenesis, Quantitative Proteomics, Labeling, CRISPR, Immunoprecipitation

Journal: Cell reports

Article Title: KRAS G12V mutation-selective requirement for ACSS2 in colorectal adenoma formation

doi: 10.1016/j.celrep.2025.115444

Figure Lengend Snippet: (A) Representative western blot for SREBP-1 and ACSS2 in mouse colon epithelial cells in response to 24-h MEK inhibition (10 nM trametinib). Results are representative of three similar experiments. (B and C) Quantification of western band intensities in (A) for cleaved SREBP-1 (B) and ACSS2 (C). One-way ANOVA; ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. (D) Intracellular cholesterol ester levels in KRAS-mutant cells in response to 24-h MEK inhibition. Data show Cholesterol Ester-Glo luminescence averages from three replicates. One-way ANOVA; ns, not significant; ** p < 0.01, **** p < 0.0001. (E) Intracellular cholesterol ester levels in KRAS mutant; Acss2 KO mouse colon epithelial cells in response to 24-h MEK inhibition. Data show Cholesterol Ester-Glo luminescence averages from three replicates. One-way ANOVA; ns, not significant. (F) Sensitivity of KRAS-mutant mouse colon epithelial cells to ACSS2 inhibition. Data show Cell-Titer Glo luminescence averages, curves were fitted with nonlinear regression in GraphPad Prism 9, and extra sum of squares f test was used to compare difference in the logarithmic value of the drug concentration that produces 50% growth inhibition (logIC50). **** p < 0.0001. (G) Sensitivity of KRAS-mutant mouse colon epithelial cells to combined MEK and ACSS2 inhibition. Data show Cell-Titer Glo luminescence averages, curves were fitted with nonlinear regression in GraphPad Prism 9, and extra sum of squares f test was used to compare difference in logIC50. **** p < 0.0001. (H) Sensitivity of KRAS-mutant, Acss2 KO mouse colon epithelial cells to MEK inhibition. Data show Cell-Titer Glo luminescence averages, curves were fitted with nonlinear regression in GraphPad Prism 9, and extra sum of squares f test was used to compare difference in logIC50. **** p < 0.0001. (I) EC50 values of trametinib sensitivity for each biological replicate, as calculated in GraphPad Prism, were plotted by mutant and ACSS2 status. One-way ANOVA; ns, not significant; *** p < 0.001.

Article Snippet: ACSS2 inhibitor , Selleck , Cat# S8588.

Techniques: Western Blot, Inhibition, Mutagenesis, Concentration Assay

Journal: Cell reports

Article Title: KRAS G12V mutation-selective requirement for ACSS2 in colorectal adenoma formation

doi: 10.1016/j.celrep.2025.115444

Figure Lengend Snippet: (A) Sensitivity of SW48 KRAS WT, G12D, and G12V isogenic cell line system to an ACSS2 inhibitor. (B) Sensitivity of LIM1215 KRAS WT, G12D, G12V, and G13D isogenic cell lines to an ACSS2 inhibitor. (C) Sensitivity of SW48 isogenic cell lines to MEK inhibition alone or in combination with an ACSS2 inhibitor. Area under the curve (AUC) was calculated in GraphPad Prism. Blue is calculated AUC in response to MEK inhibition alone; red is AUC in response to MEK and ACSS2 inhibition; *** p < 0.001, **** p < 0.0001. (D) Sensitivity of LIM1215 isogenic cell lines to MEK inhibition alone and in combination with an ACSS2 inhibitor. AUC was calculated in GraphPad Prism. Blue is calculated AUC in response to MEK inhibition alone; red is AUC in response to MEK and ACSS2 inhibition; **** p < 0.0001. (E) Relative tumor volume for mice that were allografted with KRAS G12D, KRAS G12V, KRAS G12D; ACSS2 KO, and KRAS G12V; ACSS2 KO cells. Tumor volumes were measured every 3 days for 28 days. (F) Final tumor weight at the end of the experiment. Student t test; ns, not significant; ** p < 0.01. (G and H) Relative tumor volume for mice that were allografted with (G) KRAS G12D and (H) KRAS G12V cells. Mice were dosed with 0.2 mg/kg trametinib daily by mouth (p.o.) and/or 15 mg/kg ACSS2i every 2 days intraperitoneally (i.p.).

Article Snippet: ACSS2 inhibitor , Selleck , Cat# S8588.

Techniques: Inhibition