fasn inhibitor  (TargetMol)

Journal: Microbiology Spectrum

Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection

doi: 10.1128/spectrum.01388-25

Figure Lengend Snippet: Determine the roles of FASN in BoAHV-1 productive infection in MDBK cells using a chemical inhibitor. ( A and C ) MDBK cells in six-well plates were infected with BoAHV-1 (MOI = 1) and treated either with DMSO control or with the FASN-specific inhibitor Cerulenin at the indicated concentrations. At 24 hpi, the cells were collected either to prepare cell lysates for Western blotting using an antibody against virus-associated proteins (VMRD, cat# P170703-001, 1:5,000) ( A ) or to extract DNA for subsequent qPCR analysis using gB-specific primers ( C ). ( B ) The intensity of bands for virus-associated proteins was quantified using the freeware software Image J. Changes in the levels of individual bands upon treatment with the inhibitor were calculated relative to those of the DMSO control, which was set at 100%. ( D ) Virus titers in the supernatants were determined and expressed as TCID 50 /mL. ( E ) MDBK cells in six-well plates were infected with the virus at an MOI of 1 for 24 h and treated with either DMSO control or 10 µM Cerulenin. Total RNA was then extracted from the cells for the detection of viral mRNA using RT-qPCR. Primers specific for bICP27, viral DNA polymerase, and viral protein gC were used, respectively. The data shown are means of three independent experiments, with error bars representing standard deviations. Significance was assessed by a standard t -test (ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001).

Article Snippet: FASN-specific inhibitor Cerulenin (cat# HY-A0210) was ordered from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Infection, Control, Western Blot, Virus, Software, Quantitative RT-PCR

Journal: Microbiology Spectrum

Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection

doi: 10.1128/spectrum.01388-25

Figure Lengend Snippet: Determine the roles of FASN in BoAHV-1 productive infection in Neuro-2A cells using both siRNA and chemical inhibitor. ( A ) Neuro-2A cells in six-well plates were transfected with either scrambled siRNA (150 pmol) or individual siRNAs targeting FASN (150 pmol), referred to as siRNA1, siRNA2, siRNA3, and siRNA4, respectively. At 48 h post-transfection, cell lysates were prepared and subjected to Western blot analysis to detect FASN protein levels using an antibody against FASN (Proteintech, cat# 10624-2-AP, 1:10,000). ( C ) Neuro-2A cells in six-well plates were transfected with either scrambled siRNA (150 pmol) or siRNA1 (150 pmol). At 36 h post-transfection, the cells were infected with BoAHV-1 at an MOI of 10. After 24 h of infection, the cells were collected and subjected to Western blot analysis using a monoclonal antibody against viral protein gC (VMRD, cat#F2, 1:2,000). ( E ) Neuro-2A cells in six-well plates were infected with BoAHV-1 (MOI = 10) and treated either with DMSO control or with the FASN-specific inhibitor Cerulenin at the indicated concentrations. At 24 hpi, the cell lysates were prepared and subjected to Western blotting to detect viral protein gC. ( B, D, and F ) The band intensities were quantified using the free software Image J. The intensity of each band was first normalized to that of the respective loading control Tubulin, and then normalized to that of either DMSO-treated or scrambled siRNA-transfected control, which was arbitrarily set to 100%. The data shown are means of three independent experiments, with error bars representing standard deviations. Significance was assessed using a Student’s t -test (ns, not significant; * P < 0.05; ** P < 0.01).

Article Snippet: FASN-specific inhibitor Cerulenin (cat# HY-A0210) was ordered from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Infection, Transfection, Western Blot, Control, Software

Journal: Microbiology Spectrum

Article Title: Fatty acid synthase may facilitate the trafficking of bovine alpha herpesvirus 1 out of the Golgi apparatus, potentially promoting viral infection

doi: 10.1128/spectrum.01388-25

Figure Lengend Snippet: Determine the effects of FASN inhibitor, Cerulenin, on the accumulation of gD in the Golgi apparatus. ( A ) Diagram shows the treatment manner of virus-infected cells by both Cerulenin and Monensin. ( B ) MDBK cells of either mock-infected or virus-infected were treated either with vehicle control DMSO or with Cerulenin (10 µM) or Monensin (10 µM) for a duration of 4 h prior to the termination of infection. At 24 hpi, the cells were collected to purify the Golgi apparatus using a commercial purification kit (Beijing Biolabo Technology, cat# HR0247-50T). Subsequently, Western blot analysis was performed to detect the protein levels of gD in the Golgi fractions. GOLGA1, a marker of the Golgi apparatus, was used as a loading control and for subsequent quantitative analysis. ( C ) The band intensity was analyzed with the free software Image J. The intensity of each band was first normalized to that of the respective loading control GOLGA1, and then normalized to that of the control treated with DMSO, which was arbitrarily set to 1. The data shown are means of three independent experiments with error bars indicating standard deviations. Significance was assessed with a Student’s t -test (* P < 0.05, ** P < 0.01).

Article Snippet: FASN-specific inhibitor Cerulenin (cat# HY-A0210) was ordered from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Virus, Infection, Control, Purification, Western Blot, Marker, Software

Journal: bioRxiv

Article Title: Lipodystrophy and recovery are mediated by the Wnt/lipogenesis axis during skin fibrosis

doi: 10.1101/2025.09.24.678215

Figure Lengend Snippet: A) Schematic of differentiated adipocyte cell culture model. B) Oil red O staining of undifferentiated, differentiated and 5-day 7µM WNT agonist CHIR99021 treated cells (scale bar = 50um) and respective image quantification of adipocyte lipid droplets using CellProfiler (n=6). C) Relative quantity of de-novo lipogenesis enzymes mRNA Slc2a4 ( GLUT4 ) , Acly and Fasn normalized to Hprt housekeeping gene in differentiated and WNT agonist CHIR99021 treated adipocytes (n=3). D) Band intensity and corresponding quantification of FASN protein (273 kDa) normalized to GAPDH i(n=3). E) Relative quantity of de-novo lipogenesis transcription factor Mlxipl mRNA normalized to Hprt (n=3-4). F) Number of lipid droplets per fixed field (n=8) quantified using CellPose. G) Histogram of individual lipid droplet area (in microns 2 ) (n= 1724 randomly selected droplets). H) Schematic showing experimental design of WNT agonism, FASN inhibition and 13 C acetate pulse (DNL assay) in the differentiated adipocyte cell culture model. I) Representative undifferentiated, differentiated, 5-day WNT agonist CHIR99021 treated and 5-day 250nM TVB3664 (FASN inhibitor) treated cultures co-stained with LipidSpot (lipid droplet, green) and DAPI (nucleus, blue) with inset (white square) showing high magnification single cell morphology (scale bar=100μm, for inset = 25μm) with corresponding graph quantifying the area of LipidSpot dye normalized to nuclei counts (n=8). J) Quantification of moles of neutral triglyceride (TAG) from cultures (n=4). K) Lipidomics analysis of WNT agonist BML-284 treated and FASN i treated adipocytes showing non-esterified fatty acids (NEFA) grouped by total NEFA, medium chain FA (MCFA), mono-unsaturated FA (MUFA) and poly-unsaturated FA (PUFA) normalized to protein quantity (n=4). L) 13 C-isotope incorporation in 16:0 (palmitate) species (n=4). For two groups, unpaired two-tailed Students t-test and for three or more groups, one-way ANOVA with appropriate posthoc test was employed. For the frequency distribution, non- parametric Kruskal-Wallis test was applied. *, **, ***, **** is p-value <0.05, 0.01, 0.001 and 0.0001 respectively. A p-value < 0.05 is considered significant.

Article Snippet: FASN inhibitor TVB3664 (MCE, HY-120062) was formulated in 10% DMSO and corn oil at a concentration of 0.5 mg/mL.

Techniques: Cell Culture, Staining, Inhibition, Two Tailed Test

Journal: bioRxiv

Article Title: Lipodystrophy and recovery are mediated by the Wnt/lipogenesis axis during skin fibrosis

doi: 10.1101/2025.09.24.678215

Figure Lengend Snippet: A) Schematic of regimen for FASN inhibitor treatment. During reversal, mice were given 3mg/mL TVB3664 every day for 10 days by oral gavage. B) Histology of healthy p52 mouse dorsal skin, 10-day reversal βcat istab mouse skin with vehicle gavage and FASN-inhibited 10-day reversal βcat istab mouse skin stained with Massons Trichrome (scale bar = 100μm) C) Dermal and DWAT thickness measurements as absolute values (n=5-7). D) Immunostaining for FASN (red) and Perilipin1 (green) on mouse skin. Arrows point to lipid depleted dermal adipocytes with functionally inhibited FASN protein (scale bar = 50μm). E) Histogram of adipocyte area in all three groups (n=5-6). F) Triglyceride quantity measurements normalized to weight of tissue sample G) Lipidomics analysis of non-esterified fatty acids (NEFA) grouped into by total NEFA, medium chain FA (MCFA), mono-unsaturated FA (MUFA) and poly-unsaturated FA (PUFA) normalized to weight of tissue (n=3) from whole skin of p52 control, 10-day reversal βcat istab mouse skin with vehicle gavage and FASN-inhibited 10-day reversal βcat istab (n=3). P-values were determined by Ordinary 1-way ANOVA with appropriate posthoc test. For the frequency distribution, non-parametric Kruskal-Wallis test was applied. *, **, ***, **** is p-value <0.05, 0.01, 0.001 and 0.0001 respectively. A p-value < 0.05 is considered significant.

Article Snippet: FASN inhibitor TVB3664 (MCE, HY-120062) was formulated in 10% DMSO and corn oil at a concentration of 0.5 mg/mL.

Techniques: Staining, Immunostaining, Control