Journal: Cell reports

Article Title: A multienzyme S-nitrosylation cascade regulates cholesterol homeostasis

doi: 10.1016/j.celrep.2022.111538

Figure Lengend Snippet:

Article Snippet: Rabbit polyclonal ACAT2 , Cell Signaling , 11,814.

Techniques: Recombinant, Cholesterol Assay, Enzyme-linked Immunosorbent Assay, Stable Transfection, Expressing, shRNA, Mutagenesis, Software

Journal: Lipids in Health and Disease

Article Title: Pioglitazone prevents cholesterol gallstone formation through the regulation of cholesterol homeostasis in guinea pigs with a lithogenic diet

doi: 10.1186/s12944-019-1159-4

Figure Lengend Snippet: Pioglitazone increases ABCG5 and ABCG8 and decreases NPC1L1 and ACAT2 in intestine. a Western blot analysis of ABCG5, ABCG8, NPC1L1 and ACAT2 protein; b Quantification of ABCG5 expressed as the ratio (as a percentage) of control; c Quantification of ABCG8 expressed as the ratio (as a percentage) of control; d Quantification of NPC1L1 expressed as the ratio (as a percentage) of control; e Quantification of ACAT2 expressed as the ratio (as a percentage) of control. Values shown are means ±SD, n = 4. *p < 0.05 and **p < 0.01 versus LD group

Article Snippet: The antibodies of ACAT2 (13294S) and PPARγ (2443S) were from Cell Signaling Technology Inc. (Boston, USA).

Techniques: Western Blot

Journal: Nature Communications

Article Title: Systems analysis of intracellular pH vulnerabilities for cancer therapy

doi: 10.1038/s41467-018-05261-x

Figure Lengend Snippet: Validation of systems analyses in predicting pHi-sensitive metabolic vulnerabilities. a pHi measurements of naïve and acid-adapted (AA) breast cancer MCF7 cells, under normoxia and following the inhibition of NHE1 , by cariporide treatment. For pHi measurments at least 30 cells were analyzed. b Efficiency of knockdown of indicated targets at the mRNA and protein levels, following reverse transfection of MCF7 cells with the indicated siRNAs. qPCR was repeated at least three times with three replicates. c The effect of gene inhibition in normal and low extracellular pH (pHe) shown for naïve and AA MCF7 breast cancer cells. Similar color code to Fig. is applied. At low pHe where the lowest pHi was obtained there is a large reduction in the viability of cells. In AA cells, only the selective and pH-specific targets ( GAPDH , GPI , and ACAT2 ) achieve amplified anti-proliferative effects following NHE1 inhibition, despite the smaller reduction in the pHi of these cells. PFAS , a selective but not pH-specific target, is similar to control cells following NHE1 inhibition. Knockdown of RPIA had a weak effect in naïve cells and no/opposite effects in AA cells. The viability assay was done three times with four replicates each time. The bars depict the mean and the error bars depict the standard deviation of the mean

Article Snippet: Membranes were incubated with primary antibodies against GAPDH (Cat# 2118 Cell Signaling, 1:2000), GPI (ab68643, Abcam, 1:1000), ACAT2 (Cat# 13294s Cell Signaling 1:1000), RPIA (ab181235, abcam, 1:500), PFAS (PA554628, Thermofisher, 1:200), MCT1 (sc-365501, Santa Cruz Biotechnology, 1:500), MCT4 (sc-376140, Santa Cruz Biotechnology, 1:500), and β-Actin (A5441, Sigma, 1:6000).

Techniques: Inhibition, Transfection, Amplification, Viability Assay, Standard Deviation