bms-303141  (MedChemExpress)

Journal: Neoplasia (New York, N.Y.)

Article Title: Metabolic regulation of histone acetylation by ACLY supports MDR1 expression in colorectal cancer and highlights a targetable vulnerability

doi: 10.1016/j.neo.2026.101314

Figure Lengend Snippet: ACLY activity regulates MDR1 expression in colorectal cancer. (A) Transcript levels of ACLY and MDR1 (ABCB1) in colorectal cancer (red) and normal colon tissues (grey) analyzed using GEPIA (TCGA/GTEx datasets). (B) Immunoblot analysis of MDR1 in SW480 and DLD1 cells treated with the ACLY inhibitor BMS-303141 (20 or 50 μM, 48 h). (C) Immunoblot analysis of ACLY and MDR1 in SW480 and DLD1 cells transduced with empty vector (pLV) or ACLY-overexpressing vector (pLV[Exp]-hACLY). (D) Immunoblot analysis of ACLY and MDR1 in control and ACLY-overexpressing cells treated with BMS-303141 (50 μM, 48 h). (E) Relative ABCB1 mRNA levels in control and ACLY-overexpressing cells, and in cells treated with BMS-303141. (F) Relative ACLY mRNA levels under the same conditions. Data are presented as mean ± SD (n = 3 unless otherwise indicated). Statistical significance was determined using unpaired two-tailed t-tests or one-way ANOVA with Tukey’s post hoc test. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The ACLY inhibitor BMS-303141 (MedChemExpress; HY-16107) was dissolved in DMSO (10 mM stock) and used at 50 μM for 48 h. Vorinostat (MedChemExpress; HY-10221) was dissolved in DMSO and used at 0.5 μM (SW480) or 3.5 μM (DLD1) for 48 h. Sodium ascorbate (Sigma-Aldrich; A7632) was freshly prepared in sterile water (100 mM stock) and used at 5 mM for 6 h unless otherwise indicated.

Techniques: Activity Assay, Expressing, Western Blot, Transduction, Plasmid Preparation, Control, Two Tailed Test

Journal: Neoplasia (New York, N.Y.)

Article Title: Metabolic regulation of histone acetylation by ACLY supports MDR1 expression in colorectal cancer and highlights a targetable vulnerability

doi: 10.1016/j.neo.2026.101314

Figure Lengend Snippet: ACLY activity modulates histone acetylation and MDR1 expression. (A) Immunoblot analysis of MDR1 in SW480 and DLD1 cells treated with the histone deacetylase inhibitor vorinostat (VOR; 0.5 μM for SW480 and 3.5 μM for DLD1) or DMSO for 24 h. Representative blots and densitometric quantification relative to control are shown. (B) Immunoblot analysis of acetylated histone H3 (H3K9ac) and histone H4 (H4K16ac) in SW480 wild-type (WT) and ACLY-overexpressing (OE) cells treated with vehicle or the ACLY inhibitor BMS-303141 (50 μM, 48 h). (C) Immunoblot analysis of H3K9ac and H4K16ac in DLD1 cells under the same conditions. β-actin was used as a loading control. Data are presented as mean ± SD (n = 3). Statistical significance was determined using unpaired two-tailed t-tests. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The ACLY inhibitor BMS-303141 (MedChemExpress; HY-16107) was dissolved in DMSO (10 mM stock) and used at 50 μM for 48 h. Vorinostat (MedChemExpress; HY-10221) was dissolved in DMSO and used at 0.5 μM (SW480) or 3.5 μM (DLD1) for 48 h. Sodium ascorbate (Sigma-Aldrich; A7632) was freshly prepared in sterile water (100 mM stock) and used at 5 mM for 6 h unless otherwise indicated.

Techniques: Activity Assay, Expressing, Western Blot, Histone Deacetylase Assay, Control, Two Tailed Test

Journal: Neoplasia (New York, N.Y.)

Article Title: Metabolic regulation of histone acetylation by ACLY supports MDR1 expression in colorectal cancer and highlights a targetable vulnerability

doi: 10.1016/j.neo.2026.101314

Figure Lengend Snippet: ACLY expression is associated with resistance-related transcriptional programs in colorectal cancer. (A) Correlation analysis between ACLY expression and a gene set associated with lipid metabolism (ACLY, ACSS2, ACSS1, FASN, SREBP1) and drug transport pathways (ABCB1, ABCC2, ABCG5, EpCAM, CD24) in colorectal cancer samples using GEPIA2 (TCGA dataset). (B) Schematic representation of a proposed model linking ACLY-dependent acetyl-CoA production to histone acetylation and transcriptional regulation in CRC cells. (C) Relative mRNA expression of EpCAM, ABCC2, and CD24 in SW480 and DLD1 cells overexpressing ACLY compared with empty vector controls. (D) Relative mRNA expression of EpCAM, ABCC2, and CD24 in SW480 and DLD1 cells treated with the ACLY inhibitor BMS-303141 (50 μM) compared with vehicle-treated controls. Gene expression levels were determined by qPCR and normalized to ACTB. Data are presented as mean ± SEM (n = 3). Statistical significance was determined using unpaired two-tailed t-tests. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: The ACLY inhibitor BMS-303141 (MedChemExpress; HY-16107) was dissolved in DMSO (10 mM stock) and used at 50 μM for 48 h. Vorinostat (MedChemExpress; HY-10221) was dissolved in DMSO and used at 0.5 μM (SW480) or 3.5 μM (DLD1) for 48 h. Sodium ascorbate (Sigma-Aldrich; A7632) was freshly prepared in sterile water (100 mM stock) and used at 5 mM for 6 h unless otherwise indicated.

Techniques: Expressing, Drug Transport Assay, Plasmid Preparation, Gene Expression, Two Tailed Test

Journal: NAR Molecular Medicine

Article Title: Human pneumovirus induces IFN-dependent expression of the immune-responsive gene 1 and is inhibited by 4-octyl itaconate in human macrophages

doi: 10.1093/narmme/ugag017

Figure Lengend Snippet: 4OI reduces expression of ATP-dependent citrate lyase required for HMPV replication. ( A ) 4OI reduces ATP-dependent citrate lyase ( ACLY) expression. MDMs were treated with 4OI (250 µM) or DMSO prior to infection with HMPV for 24 h. Expression levels of ACLY, FASN , and SCD1 mRNA were quantified relative to untreated, uninfected MDMs via qRT-PCR ( n ≥ 4). ( B ) ACLY inhibition reduces HMPV levels. MDMs were pretreated with 10 or 20 µM of the ACLY inhibitor BMS-303141 or DMSO prior to infection with HMPV for 24 h. HMPV N-gene mRNA was determined by qRT-PCR ( n = 4). Protein expression of HMPV N and GAPDH was analysed via immunoblotting of whole cell lysates ( n = 2). Protein levels were quantified by normalizing of band intensities against GAPDH and expressed as fold change compared to siNTC-transfected, infected MDMs. Multiple comparisons were analysed by a paired one-way ANOVA with Tukey post-hoc test. Significance was ranked as * P < .05; ** P < .01; *** P < .001; **** P < .0001 and ns, not significant. ( C ) HMPV stimulates increased level of neutral lipids in infected MDMs. MDMs were left uninfected (−) or infected with a MOI 1 of GFP-expressing recombinant HMPV for 24 h. HCS LipidTOX™ Deep Red neutral lipid stain was used to monitor lipids by confocal microscopy. Left panels: Representative images for each treatment showing HCS LipidTOX™ Deep Red (magenta) and HMPV (green). Right panel: Quantification of the LipidTOX™ signal per area. Signal was quantified from z-stacks of six fields of view per condition using the 20× numerical aperture yielding to ~2000 cells per condition and normalized to the area. Scalebar is adjusted to 100 µm ( n = 1). The data were assessed for normality and then compared with a non-parametric, two-tailed Mann–Whitney test. Significance was ranked as * P < .05; ** P < .01; *** P < .001; **** P < .0001, and ns, not significant.

Article Snippet: Inhibition with S-Ruxolitinib (Merck) or GSK 872 (GlaxoSmithKline) was performed 1 h before infection with HMPV, whereas BMS-303141 (MedChemExpress) was preincubated for 2 h. The IFNAR neutralizing antibody (nIFNAR, clone MMHAR-2) was purchased from PBL; 5 or 10 μg/ml were incubated for 30 min before infection with HMPV.

Techniques: Expressing, Infection, Quantitative RT-PCR, Inhibition, Western Blot, Transfection, Recombinant, Staining, Confocal Microscopy, Two Tailed Test, MANN-WHITNEY

Journal: NAR Molecular Medicine

Article Title: Human pneumovirus induces IFN-dependent expression of the immune-responsive gene 1 and is inhibited by 4-octyl itaconate in human macrophages

doi: 10.1093/narmme/ugag017

Figure Lengend Snippet: Exogenously added 4OI, but not itaconate or citraconate, reduces HMPV levels in human MDMs. (A–C) MDMs were preincubated with 100 or 250 µM 4OI or its vehicle, DMSO, prior to HMPV infection for 24 h. Levels of HMPV N-gene mRNA were quantified by qRT-PCR ( A ; n ≥ 5), HMPV N and GAPDH protein levels were analysed by immunoblotting ( B ; n ≥ 5), while infectious HMPV/HMPV titers in MDM supernatants was determined as TCID 50 /ml ( C ; n = 5). (D, E) MDMs were treated with itaconate (ITA; 0.5, 5, 10, and 20 mM) prior to HMPV infection for 24 h. Levels of HMPV N-gene mRNA were quantified by qRT-PCR ( D ; n ≥ 2), while HMPV N and GAPDH proteins were analysed by immunoblotting ( E ; n ≥ 2). (F, G) MDMs were treated with citraconate (CIT; 1, 10, 20, and 50 mM) prior to HMPV infection for 24 h. Levels of the HMPV N-gene mRNA were quantified by qRT-PCR ( F ; n = 4), while HMPV N and GAPDH protein levels were analysed by immunoblotting ( G ; n = 2). Protein levels were quantified by normalizing band intensities against GAPDH and expressed as fold change compared to untreated (i.e. no itaconate variant) but infected MDMs. Multiple comparisons were analysed by a paired onw-way ANOVA with Tukey post-hoc test. Significance was ranked as * P < .05; ** P < .01; *** P < .001; **** P < .0001, and ns, not significant.

Article Snippet: Inhibition with S-Ruxolitinib (Merck) or GSK 872 (GlaxoSmithKline) was performed 1 h before infection with HMPV, whereas BMS-303141 (MedChemExpress) was preincubated for 2 h. The IFNAR neutralizing antibody (nIFNAR, clone MMHAR-2) was purchased from PBL; 5 or 10 μg/ml were incubated for 30 min before infection with HMPV.

Techniques: Infection, Quantitative RT-PCR, Western Blot, Variant Assay

Journal: NAR Molecular Medicine

Article Title: Human pneumovirus induces IFN-dependent expression of the immune-responsive gene 1 and is inhibited by 4-octyl itaconate in human macrophages

doi: 10.1093/narmme/ugag017

Figure Lengend Snippet: The Nrf2 pathway is induced by 4OI, but not by itaconate or citraconate, and limits HMPV in human macrophages. MDMs were preincubated with ( A ) 100 and 250 µM of 4OI or DMSO, ( B ) 0.5, 5, 10, and 20 mM of itaconate, and ( C ) 1, 10, 20, and 50 mM of citraconate prior to infection with HMPV for 24 h. (A–C) Levels of HO1 and NQO1 mRNA were quantified by qRT-PCR. The expression of HO1 and GAPDH protein was analysed using immunoblotting of whole cell lysates. ( A ) n ≥ 5 for mRNA and protein; ( B ) n ≥ 2 for mRNA and protein; ( C ) n = 4 for mRNA and n = 2 for protein. HO1 and NQO1 mRNA levels were assessed relative to untreated, uninfected MDMs for all experiments. HO1 protein levels were quantified by normalization of band intensities against GAPDH and expressed as fold change compared to untreated, infected MDMs. ( D ) MDMs were transfected with 20 nM NFE2L2 (Nrf2) siRNA or siNTC and infected with HMPV for 24 h. Nrf2, HMPV N protein, and GAPDH were analysed by immunoblotting of whole cell lysates and quantified ( n = 4). Nrf2 protein level is presented as fold change relative to siNTC-treated uninfected MDMs (left panel), while HMPV N-protein is presented as relative to siNTC-treated HMPV-infected MDMs (right panel). In panels (A–C), multiple comparisons were analysed by a paired onw-way ANOVA with Tukey post-hoc test, in panel (D) unpaired Student’s t - test. Significance was ranked as * P < .05; ** P < .01; *** P < .001; **** P < .0001 and ns, not significant.

Article Snippet: Inhibition with S-Ruxolitinib (Merck) or GSK 872 (GlaxoSmithKline) was performed 1 h before infection with HMPV, whereas BMS-303141 (MedChemExpress) was preincubated for 2 h. The IFNAR neutralizing antibody (nIFNAR, clone MMHAR-2) was purchased from PBL; 5 or 10 μg/ml were incubated for 30 min before infection with HMPV.

Techniques: Infection, Quantitative RT-PCR, Expressing, Western Blot, Transfection

Journal: NAR Molecular Medicine

Article Title: Human pneumovirus induces IFN-dependent expression of the immune-responsive gene 1 and is inhibited by 4-octyl itaconate in human macrophages

doi: 10.1093/narmme/ugag017

Figure Lengend Snippet: HMPV induction of IRG1 in human macrophages is dependent on type I IFN. ( A ) MDMs were infected with HMPV for the indicated time points. HMPV N gene or IRG1 mRNA levels were determined by qRT-PCR analysis ( n = 3), while protein levels of IRG1, HMPV N, and GAPDH were analysed by immunoblotting ( n = 2). IRG1 protein levels shown are for the same donor sample used in Fig. of , thus panels showing HMPV N and GAPDH levels being identical to the ones shown . ( B ) MDMs were treated with 500 ng/ml LPS for 2 h. IRG1 mRNA levels were assessed by qRT-PCR and normalized to untreated (−) control ( n = 5). ( C ) MDMs were transfected with 10 μg/ml poly IC for 6 or 24 h. IRG1 mRNA levels were assessed by qRT-PCR and normalized to untreated (−) cells ( n ≥ 2). ( D ) MDMs were pre-incubated (30 min) or not with 10 μg/ml of neutralizing IFNAR antibody before infection with HMPV for 24 h. IRG1 mRNA expression was determined using qRT-PCR and normalized to untreated (−) cells ( n = 4). ( E ) MDMs were treated with 100 U/ml recIFN-β for 24 h. IRG1 mRNA expression was determined using qRT-PCR and normalized to untreated (−) cells ( n = 4). ( F ) MDMs were preincubated with the JAK1/JAK2 inhibitor ruxolitinib (5 or 10 μM) prior to infection with HMPV for 18 h or treatment with IFN-β for 3 h. Protein levels in whole cell lysates were analysed for IRG1, STAT1 (Tyr701), STAT1, and GAPDH via immunoblot ( n = 2).

Article Snippet: Inhibition with S-Ruxolitinib (Merck) or GSK 872 (GlaxoSmithKline) was performed 1 h before infection with HMPV, whereas BMS-303141 (MedChemExpress) was preincubated for 2 h. The IFNAR neutralizing antibody (nIFNAR, clone MMHAR-2) was purchased from PBL; 5 or 10 μg/ml were incubated for 30 min before infection with HMPV.

Techniques: Infection, Quantitative RT-PCR, Western Blot, Control, Transfection, Incubation, Expressing

Journal: NAR Molecular Medicine

Article Title: Human pneumovirus induces IFN-dependent expression of the immune-responsive gene 1 and is inhibited by 4-octyl itaconate in human macrophages

doi: 10.1093/narmme/ugag017

Figure Lengend Snippet: TBK1 and NF-κB positively regulate IRG1 levels in HMPV-infected human macrophages. MDMs were incubated with 5 or 10 μM of the TBK1 inhibitor BX795 for 30 min before infection with HMPV for 24 h. ( A ) IRG1 and IFN-β mRNA levels ( n ≥ 3) were analysed by qRT-PCR (left panels), while in panel ( B ) IRG1 and GAPDH protein levels were determined by immunoblotting, quantified, and presented with SD relative to uninfected cells treated with DMSO ( n = 2; right panel). (C–K) MDMs were transfected with siRNAs targeting the NF-κB subunit RELA (C–E), IRF1 (F–H), RIPK3 (I–K), or control siRNA (siNTC) before infection with HMPV for 6, 9, or 24 h and analysis of IRG1 mRNA levels by qRT-PCR or IRG1, RelA/p65, IRF1, RIPK3, or GAPDH protein levels by immunoblotting. IRG1 mRNA levels were assessed relative to siNTC-transfected, uninfected MDMs for all experiments. ( C, E ) IRG1 mRNA after 6 h ( n = 3) or 24 h HMPV ( n = 3). ( D ) Protein levels of IRG1, RelA/p65, and GAPDH ( n = 3) after 9 h HMPV. ( F, H ) IRG1 mRNA after 6 h ( n = 4) or 24 h of infection ( n = 3). ( G ) Protein levels of IRG1, IRF1, and GAPDH ( n = 3) after 9 h HMPV. ( I, K ) IRG1 mRNA after 6 h ( n = 3) or 24 h of infection ( n = 3). ( J ) Protein levels of IRG1, IRF1, and GAPDH ( n = 3) after 9 h HMPV. ( L ) MDMs were incubated with 100 nM of the RIPK3 inhibitor GSK 872 for 1 h before infection with HMPV for 24 h. IRG1 mRNA levels were normalized relative to uninfected cells treated with DMSO ( n = 2). Single comparison between control (NTC) siRNA and target-siRNA conditions were calculated using paired t -test with Tukey post-hoc test. * P < .05, ** P < .01, *** P < .001, **** P < .0001; ns = non-significant. See also

Article Snippet: Inhibition with S-Ruxolitinib (Merck) or GSK 872 (GlaxoSmithKline) was performed 1 h before infection with HMPV, whereas BMS-303141 (MedChemExpress) was preincubated for 2 h. The IFNAR neutralizing antibody (nIFNAR, clone MMHAR-2) was purchased from PBL; 5 or 10 μg/ml were incubated for 30 min before infection with HMPV.

Techniques: Infection, Incubation, Quantitative RT-PCR, Western Blot, Transfection, Control, Comparison

Journal: NAR Molecular Medicine

Article Title: Human pneumovirus induces IFN-dependent expression of the immune-responsive gene 1 and is inhibited by 4-octyl itaconate in human macrophages

doi: 10.1093/narmme/ugag017

Figure Lengend Snippet: 4OI, itaconate, and citraconate differ in regulating IRG1 and the IFN-β response upon HMPV infection. MDMs were preincubated with ( A ) 100 and 250 µM of 4OI or DMSO, ( B ) 0.5, 5, 10, and 20 mM of itaconate, and ( C ) 1, 10, 20, and 50 mM of citraconate prior to infection with HMPV for 24 h. Levels of IRG1 and IFNB mRNA (left panels) were quantified by qRT-PCR. The expression of STAT1 (Tyr701), STAT1, IRG1, and GAPDH protein (right panels) was analysed using immunoblotting of whole-cell lysates and quantification STAT1 (Tyr701) expression. ( A ) n ≥ 5 for mRNA and protein; ( B ) n ≥ 2 for mRNA and protein; and ( C ) n = 4 for mRNA and n = 2 for protein. IFNB and IRG1 mRNA levels were assessed relative to untreated, uninfected MDMs for all experiments. Protein levels were quantified by normalization of band intensities against GAPDH and was expressed as fold change compared to untreated, uninfected MDMs. Multiple comparisons were analysed by a paired onw-way ANOVA with Tukey post-hoc test. Significance was ranked as * P < .05; ** P < .01; *** P < .001; **** P < .0001, and ns, not significant.

Article Snippet: Inhibition with S-Ruxolitinib (Merck) or GSK 872 (GlaxoSmithKline) was performed 1 h before infection with HMPV, whereas BMS-303141 (MedChemExpress) was preincubated for 2 h. The IFNAR neutralizing antibody (nIFNAR, clone MMHAR-2) was purchased from PBL; 5 or 10 μg/ml were incubated for 30 min before infection with HMPV.

Techniques: Infection, Quantitative RT-PCR, Expressing, Western Blot

Journal: NAR Molecular Medicine

Article Title: Human pneumovirus induces IFN-dependent expression of the immune-responsive gene 1 and is inhibited by 4-octyl itaconate in human macrophages

doi: 10.1093/narmme/ugag017

Figure Lengend Snippet: 4OI reduces expression of ATP-dependent citrate lyase required for HMPV replication. ( A ) 4OI reduces ATP-dependent citrate lyase ( ACLY) expression. MDMs were treated with 4OI (250 µM) or DMSO prior to infection with HMPV for 24 h. Expression levels of ACLY, FASN , and SCD1 mRNA were quantified relative to untreated, uninfected MDMs via qRT-PCR ( n ≥ 4). ( B ) ACLY inhibition reduces HMPV levels. MDMs were pretreated with 10 or 20 µM of the ACLY inhibitor BMS-303141 or DMSO prior to infection with HMPV for 24 h. HMPV N-gene mRNA was determined by qRT-PCR ( n = 4). Protein expression of HMPV N and GAPDH was analysed via immunoblotting of whole cell lysates ( n = 2). Protein levels were quantified by normalizing of band intensities against GAPDH and expressed as fold change compared to siNTC-transfected, infected MDMs. Multiple comparisons were analysed by a paired one-way ANOVA with Tukey post-hoc test. Significance was ranked as * P < .05; ** P < .01; *** P < .001; **** P < .0001 and ns, not significant. ( C ) HMPV stimulates increased level of neutral lipids in infected MDMs. MDMs were left uninfected (−) or infected with a MOI 1 of GFP-expressing recombinant HMPV for 24 h. HCS LipidTOX™ Deep Red neutral lipid stain was used to monitor lipids by confocal microscopy. Left panels: Representative images for each treatment showing HCS LipidTOX™ Deep Red (magenta) and HMPV (green). Right panel: Quantification of the LipidTOX™ signal per area. Signal was quantified from z-stacks of six fields of view per condition using the 20× numerical aperture yielding to ~2000 cells per condition and normalized to the area. Scalebar is adjusted to 100 µm ( n = 1). The data were assessed for normality and then compared with a non-parametric, two-tailed Mann–Whitney test. Significance was ranked as * P < .05; ** P < .01; *** P < .001; **** P < .0001, and ns, not significant.

Article Snippet: Inhibition with S-Ruxolitinib (Merck) or GSK 872 (GlaxoSmithKline) was performed 1 h before infection with HMPV, whereas BMS-303141 (MedChemExpress) was preincubated for 2 h. The IFNAR neutralizing antibody (nIFNAR, clone MMHAR-2) was purchased from PBL; 5 or 10 μg/ml were incubated for 30 min before infection with HMPV.

Techniques: Expressing, Infection, Quantitative RT-PCR, Inhibition, Western Blot, Transfection, Recombinant, Staining, Confocal Microscopy, Two Tailed Test, MANN-WHITNEY