acly inhibitor bms 303141  (MedChemExpress)

Journal: bioRxiv

Article Title: Ratiometric Fluorescent Protein Biosensors Reveal Citrate Dynamics and Cellular Heterogeneity

doi: 10.64898/2026.04.16.718871

Figure Lengend Snippet: A-B. Pseudo-colored images of HeLa cells expressing ratioCitron1Low ( A ) and ratioCitron1High ( B ) in cytosol before and after addition of citrate. Scale bar: 100 µm. C-D. Δ R / R and Δ F / F values of ratioCitron1Low ( C ) and ratioCitron1High ( D ) versus time upon the addition of citrate. HeLa cells were incubated with 4 μM digitonin for 10 mins before the experiment and a final concentration of 20 mM citrate was added at t = 0 (ratioCitron1Low n = 16, ratioCitron1High n = 19, mean ± SD). E. Time courses of Δ R / R ₀ of ratioCitron1 variants (ratioCitron1Low ( n = 20), ratioCitron1High ( n = 10), ratioCitron1Low-con ( n = 5), ratioCitron1High-con ( n = 5)) expressed in HeLa cells upon the citrate titration. HeLa cells expressing ratioCitron1 variants were treated with digitonin and citrate titration was carried out. The final concentration of citrate is 100 μM at 0 min, 1 mM at 5 min, 10 mM at 10 min (mean ± SD). F-G. Δ R / R values of ratioCitron1 variants expressed in cytosol ( F ) and in mitochondria (mito-ratioCiton1) ( G ) of HeLa cells upon the addition of the ACLY inhibitor of BMS-303141. H. Δ R / R values of ratioCitron1 variants expressed in mitochondria (mito-ratioCitron1) of HeLa cells upon the addition of MPC inhibitor UK-5099 (final concentration: 25 μM). Δ F/F = ( F on - F off )/ F off , and F with excitation 405 nm or 470 nm and emission 518/45 nm. Δ R / R = ( R on - R off )/ R off , and R = ( F with excitation 470 nm and emission 518/45 nm) / ( F with excitation 405 nm and emission 518/45 nm).

Article Snippet: For imaging the treatment with MPC inhibitor UK-5099 (MedChemExpress) and ACLY inhibitor BMS-303141 (MedChemExpress), Hank’s balanced salt solution (HBSS; Nacalai Tesque, 09735-75) and 10 mM HEPES (Nacalai Tesque, 177557-94) was used as imaging buffer.

Techniques: Expressing, Incubation, Concentration Assay, Titration

Journal: bioRxiv

Article Title: A palmitate-rich metastatic niche enables metastasis growth via p65 acetylation

doi: 10.1101/2022.10.24.513556

Figure Lengend Snippet: A . Intracellular levels of acetyl-CoA in 4T1 and MCF10A H-Ras V12 cells incubated in 2D monolayer and 3D spheroids cultures for 5 days in medium containing extra palmitate. Intracellular acetyl-CoA measurements in additional cell lines (EMT6.5 and MCF7) are shown in . Two-tailed unpaired student’s T-test (n = 4). B . Relatives changes in acetyl-CoA abundance in EMT6.5 (m.f.) breast primary tumors and lung metastases. Data are shown as fold changes compared with the acetyl-CoA abundance in the primary tumors. Two-tailed unpaired student’s T-test (n = 4). C . Relatives changes in acetyl-CoA abundance in 4T1 (m.f.) breast primary tumors and lung metastases upon acute inhibition of CPT1A using the inhibitor etomoxir (30 mg/kg i.p.) or vehicle (water). Data are shown as fold changes compared with the acetyl-CoA abundance in the primary tumors of the group of mice treated with the vehicle. Data points represented as zero were below the detection limit. One-way ANOVA with Dunnett’s multiple comparison test (n ≥4). D . Relatives changes in acetyl-CoA abundance in mouse 4T1 breast cancer spheroids upon treatment with the CPT1A inhibitor etomoxir (50 μM), transduced with a lentiviral vector with shRNA against Cpt1a (knockdown), sgRNA Cpt1a (knockout) or scrambled control sequences in the presence of extra palmitate. Intracellular acetyl-CoA measurements in additional human cell lines (MCF10A H-Ras V12 and MCF7) are shown in . One-way ANOVA with Dunnett’s multiple comparison test or two-tailed unpaired student’s T-test are shown (n = 4). E and F . 3D spheroids growth upon genetic inhibition of either Cpt1a or CPT1A together with pharmacologic ALCY inhibition using BMS-303141 (20 μM) in 4T1 ( A .) and MCF10A H-Ras V12 ( B ) cells with or without extra palmitate and in the presence of the acetate as metabolic rescue (5 mM). 3D spheroid growth is represented by the average spheroids area of >100 spheroids. One-way ANOVA with Turkey’s multiple comparison test (n ≥4).

Article Snippet: The ATP Citrate lyase (ACLY) inhibitor BMS-303141 was purchased from MedChemExpress, dissolved in DMSO and used at 20 μM.

Techniques: Incubation, Two Tailed Test, Inhibition, Comparison, Transduction, Plasmid Preparation, shRNA, Knock-Out