mpc inhibitor uk 5099  (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: The FASEB Journal

Article Title: Pyruvate prevents the onset of the cachectic features and metabolic alterations in myotubes downregulating STAT3 signaling

doi: 10.1096/fj.202200848r

Figure Lengend Snippet: FIGURE 5 Myotubes treated with the MPC inhibitor UK5099 acquire cachectic features. Myotubes have been treated with UK5099 (10 μM final) for 24 h. (A) Representative images of control and UK5099-treated myotubes. (B) Myotube width obtained by ImageJ and calculated in at least 10 randomly chosen fields. (C) Ubiquitin immunoblot. (D) LC3II immunoblot. In (C) and (D) the bar graph reports the mean value of each sample obtained by the ratio with the PVDF membrane used for normalization. (E) Lactate assay. (F) Oxygen consumption rate (OCR). (G) Analysis of mitochondrial membrane potential by confocal microscopy. Mitochondria are stained with TMRM probe (red fluorescence), while blue fluorescence shows the nuclei. The bar graph reports the mean value of fluorescence for each sample measured using ImageJ software in at least 10 randomly chosen fields. (H) PDH activity. C, control myotubes. n = 3; *p < .05.

Article Snippet: Unless otherwise specified, all used reagents were obtained from Sigma- Aldrich, Inc. (St. Louis, MO, USA); SDS- PAGE materials and ECL detection reagents were purchased from Bio- Rad Laboratories, (Hercules, USA); anti- Fbx32/ Atrogin1 (ab168372), anti- OXPHOS (ab110413), antiSTAT3 (ab68153), and anti- Myosin Heavy Chain (MHC) (ab 91506) primary antibodies were from Abcam (Cambridge, UK); anti- PDH- E1 (sc- 377092) and anti- ubiquitin (sc8017) primary antibodies, mitochondrial pyruvate carrier (MPC) inhibitor UK5099 (sc- 361394) and STAT3 inhibitor WP1066 (sc- 203282) were from Santa Cruz Biotechnology (Dallas, TX, USA); anti- LC3B (#3868) and anti- phosphoTyr705- STAT3 (#9145) primary antibodies were from Cell Signaling Technology Inc. (Danvers, MA,USA); Tetramethyl- rhodamine methyl ester (TMRM) probe was from Molecular Probe (Eugene, OR, USA); K- LATE kit for lactate assay was from Megazyme (Bray, Ireland); pyridine and N- tert- Butyldimethylsilyl- N- methyltrifluoroacetamide with 1% tert- Butyldimethylchlorosilane (MBTSTFA + 1% TBDMCS) were from Pierce, ThermoFisher Scientific (Waltham, MA, USA).

Techniques: Control, Ubiquitin Proteomics, Western Blot, Membrane, Lactate Assay, Confocal Microscopy, Staining, Fluorescence, Software, Activity Assay

Journal: Oncology Research

Article Title: Lactate Maintains BCR/Abl Expression and Signaling in Chronic Myeloid Leukemia Cells Under Nutrient Restriction

doi: 10.3727/096504022X16442289212164

Figure Lengend Snippet: Effects of mono-carbon transporter 1 (MCT-1) or mitochondrial pyruvate carrier (MPC) inhibition on BCR/Abl expression/signaling and cell growth in the presence of lactate. K562 (A, C) or KCL22 (B, D) cells were cultured for 14 days as described in the legend of , in the presence or absence of the MCT-1 inhibitor AR-C155858 (A, B) or the MPC inhibitor UK5099 (C, D) from time 0 of incubation. Left panels: Histograms represent counts of trypan blue-negative cells and are means ± SEM of data obtained from three independent experiments; the statistical significance of differences was determined by the Student’s t -test for paired samples; * p < 0.05; ** p < 0.01; **** p < 0.001; horizontal lines indicate time 0 values. Right panels: Cells were lysed in Laemmli buffer and total cell lysates subjected to SDS-PAGE and immunoblotting with antibodies raised against c-Abl or p-Crkl; anti-vinculin antibody was used to verify the equalization of protein loading; one representative experiment out of three with similar outcome is shown.

Article Snippet: In some experiments, cultures were treated with the mono-carbon transporter 1/2 (MCT-1/2) inhibitor AR-C155858 (MedChemExpress, Monmouth Junction, NJ, USA) at 5 μM final concentration, or with the mitochondrial pyruvate carrier (MPC) inhibitor UK5099 (MedChemExpress) at 5 μM final concentration, or with methyl-pyruvate (Sigma-Aldrich) at 10 mM final concentration.

Techniques: Inhibition, Expressing, Cell Culture, Incubation, SDS Page, Western Blot

Journal: Cancers

Article Title: ARRB1 Regulates Metabolic Reprogramming to Promote Glycolysis in Stem Cell-Like Bladder Cancer Cells

doi: 10.3390/cancers13081809

Figure Lengend Snippet: ARRB1 regulates mitochondrial translocation of pyruvate and glucose uptake: ( a ) immunoblot for detection of glycolytic markers, MPC1 and GLUT1 in bladder cancer cell lines (253J, HT1376, 5637) and nonmalignant cells (UROTSA), as well as ARRB1-depleted HT1376 cells (KO cl. 9 and cl. 10). ( b ) Mitochondria of ARRB1-depleted HT1376 cells (KO cl. 9) and corresponding controls (NT) were collected, and mitochondria/total protein ratio was calculated. Data: means ± SEM, n = 2. The experiment was repeated once ( c ) Mitochondrial and cytoplasmic extracts were collected for colorimetric detection of pyruvate. Data: means ± SEM, n = 3; *** p < 0.001 (two-tailed unpaired t test). The experiment was repeated once ( d ) Secreted Lactate (glycolysis) in presence or absence of the MPC1-inhibitor UK-5099. Data: means ± SEM, n = 4; *** p < 0.001 (two-way ANOVA). ( e ) Glucose uptake in ARRB1-depleted HT1376 cells (KO cl. 9 and cl.10) compared to corresponding controls (NT). Data: means ± SEM, n = 4; *** p < 0.001, * p < 0.05 (One-way ANOVA). ( f ) Measurement of mitochondrial fuel usage dependency of ARRB1-depleted HT1376 cells to oxidize glucose/pyruvate, glutamine/glutamate, and long-chain fatty acids. Data: means ± SEM, n = 4 to 8; * p < 0.05, (two-way ANOVA). NS: not significant.

Article Snippet: The Seahorse cartridge was humidified with sterile water overnight in a non-CO 2 incubator at 37 °C and then submerged in prewarmed XF-calibrant solution for 45–60 min. BPTES (inhibitor of the glutaminase/glutamine oxidation pathway), Etomoxir Sodium Salt hydrate (inhibitor of carnitine palmitoyl-transferase 1 a/ long-chain fatty acid oxidation), and UK-5099 (inhibitor of mitochondrial pyruvate carrier (MPC)/ glucose oxidation pathway) were purchased from Sigma (Cat # E1905, SML0601, and PZ0160).

Techniques: Translocation Assay, Western Blot, Two Tailed Test

Journal: Molecular Metabolism

Article Title: The mitochondrial pyruvate carrier mediates high fat diet-induced increases in hepatic TCA cycle capacity

doi: 10.1016/j.molmet.2017.09.002

Figure Lengend Snippet: MPC inhibition attenuates inflammatory response to palmitate plus LPS. (A–E) Primary hepatocytes transcript abundance of (A) Il1b , (B) Ccl2 , (C) Tnfa , and (D) Tgfb after 12hr treatment with 300 μM palmitate, LPS (0.01, 0.1, 1.0 ng/mL), and UK5099 (5 μM) or vehicle control. (Hepatocytes were pooled from three mice in technical triplicate). (Data are presented as mean ± SEM; n = 3, *p < 0.05, **p < 0.01, ***p < 0.001).

Article Snippet: The following day, hepatocytes were treated with 300 μM palmitate conjugated to BSA, lipopolysaccharides (LPS) from Escherichia coli O111:B4 (Sigma, L2630), and 5 μM selective MPC inhibitor UK5099 (Tocris, #4186) in DMSO (<0.01% v/v final) or appropriate vehicle controls for 12 h. Following treatment, cells were washed twice with ice-cold PBS, lysed in TRIzol (ThermoFisher/Ambion), and snap frozen in liquid nitrogen for later processing.

Techniques: Inhibition, Control