Journal: Veterinary Research

Article Title: Mechanistic role of lipid metabolism in foot-and-mouth disease virus (FMDV) replication

doi: 10.1186/s13567-026-01762-6

Figure Lengend Snippet: The effect of ACC on FMDV replication. A Research pathway map for de novo fatty acid synthesis. B BHK-21 cells were treated with 30 μM TOFA, 60 μM malonyl‑CoA, or 30 μM TOFA plus 60 μM malonyl‑CoA. After fixation, cells were stained with Nile red, and nuclei were labeled with DAPI (blue). Scale bar, 20 µm. C BHK-21 cells were treated with 5–45 μM TOFA for 24 h, and cell viability was not significantly affected. D Cytopathic effect (CPE) following FMDV infection in the presence of increasing concentrations of TOFA. E RT–qPCR confirmed that TOFA treatment inhibits FMDV replication. F Western blot analysis confirmed that TOFA treatment suppresses FMDV replication. G TOFA inhibited FMDV replication by 50% (EC 50 ) at 21.33 μM. H CP640186 inhibited FMDV replication by 50% (EC 50 ) at 0.2021 μM. I Viral plaque assay demonstrated that TOFA treatment significantly reduces FMDV replication. J BHK-21 cells were treated with 5–120 μM malonyl‑CoA for 24 h, and cell viability was not substantially affected. K RT–qPCR showed that, in the presence of 15 μM TOFA, supplementation with 0–45 μM malonyl‑CoA partially restored FMDV replication. L Western blot analysis confirmed that, in the presence of 45 μM TOFA, addition of 60 μM malonyl‑CoA partially rescued the expression of FMDV VP1 protein.

Article Snippet: The hamster kidney fibroblasts (BHK-21) used in the experiments were purchased from Wuhan Punosai Life Science and Technology Co. Acetyl-CoA carboxylase inhibitor TOFA (HY-100568) was purchased from MedChemExpress (MCE); fatty acid synthase inhibitor C75 (HY-12364) was purchased from MCE; malonyl-CoA (HY-115899) was purchased from MCE; palmitic acid (HY-N0830) was purchased from MCE; DGAT-1 inhibitor A922500 (HY-10038) was purchased from MCE; oleic acid (HY-N1446) was purchased from MCE; CPT1A inhibitors Etomoxir (HY-50202) and CP640186 (HY-15259) were purchased from MCE; and rabbit polyclonal antibody against FMDV VP1 protein (type O) (bs-41049R) was purchased from Beijing Biosynthesis Biotechnology Co., Ltd. (bioss).

Techniques: Staining, Labeling, Infection, Quantitative RT-PCR, Western Blot, Viral Plaque Assay, Expressing

Journal: Veterinary Research

Article Title: Mechanistic role of lipid metabolism in foot-and-mouth disease virus (FMDV) replication

doi: 10.1186/s13567-026-01762-6

Figure Lengend Snippet: The effect of FASN on FMDV replication. A BHK-21 cells were treated with 30 μM C75, 75 μM palmitic acid, or 30 μM C75 plus 75 μM palmitic acid. After fixation, cells were stained with Nile red. Scale bar, 20 µm. B BHK-21 cells were treated with 5–45 μM C75 for 24 h, and cell viability was not significantly affected. C Cytopathic effect (CPE) following FMDV infection in the presence of increasing concentrations of C75. D RT–qPCR confirmed that C75 treatment inhibits FMDV replication. E Western blot analysis verified that C75 treatment suppresses the expression of FMDV VP1 protein. F Combined treatment with 10 μM C75 and TOFA exerted a stronger inhibitory effect on FMDV replication. G Viral plaque assay demonstrated that C75 treatment significantly inhibits FMDV replication. H BHK-21 cells were treated with 5–75 μM palmitic acid for 24 h, and cell viability was not substantially affected. I RT–qPCR showed that, in the presence of 30 μM C75, supplementation with 0–75 μM palmitic acid partially restored FMDV replication. J Western blot analysis confirmed that, in the presence of 45 μM C75, addition of 75 μM palmitic acid partially rescued FMDV VP1 protein expression. K BHK-21 cells were treated with 5–30 μM 2-bromopalmitate (2‑BP) for 24 h, and cell viability was not significantly affected. L RT–qPCR confirmed that 2‑BP treatment inhibits FMDV replication. M Western blot analysis verified that 2‑BP treatment suppresses the expression of FMDV VP1 protein.

Article Snippet: The hamster kidney fibroblasts (BHK-21) used in the experiments were purchased from Wuhan Punosai Life Science and Technology Co. Acetyl-CoA carboxylase inhibitor TOFA (HY-100568) was purchased from MedChemExpress (MCE); fatty acid synthase inhibitor C75 (HY-12364) was purchased from MCE; malonyl-CoA (HY-115899) was purchased from MCE; palmitic acid (HY-N0830) was purchased from MCE; DGAT-1 inhibitor A922500 (HY-10038) was purchased from MCE; oleic acid (HY-N1446) was purchased from MCE; CPT1A inhibitors Etomoxir (HY-50202) and CP640186 (HY-15259) were purchased from MCE; and rabbit polyclonal antibody against FMDV VP1 protein (type O) (bs-41049R) was purchased from Beijing Biosynthesis Biotechnology Co., Ltd. (bioss).

Techniques: Staining, Infection, Quantitative RT-PCR, Western Blot, Expressing, Viral Plaque Assay

Journal: Veterinary Research

Article Title: Mechanistic role of lipid metabolism in foot-and-mouth disease virus (FMDV) replication

doi: 10.1186/s13567-026-01762-6

Figure Lengend Snippet: Utilization of the lipid metabolism model during FMDV infection. ACC catalyzes the carboxylation of acetyl-CoA to generate malonyl-CoA; malonyl-CoA and acetyl-CoA are catalyzed by FASN to produce fatty acids; fatty acids are catalyzed by DGAT-1 to form triglycerides, which are stored in LDs; the degradation of LDs generates a large amount of FFAs, which are transported into mitochondria under the action of CPT1A to participate in β-oxidation for ATP production.

Article Snippet: The hamster kidney fibroblasts (BHK-21) used in the experiments were purchased from Wuhan Punosai Life Science and Technology Co. Acetyl-CoA carboxylase inhibitor TOFA (HY-100568) was purchased from MedChemExpress (MCE); fatty acid synthase inhibitor C75 (HY-12364) was purchased from MCE; malonyl-CoA (HY-115899) was purchased from MCE; palmitic acid (HY-N0830) was purchased from MCE; DGAT-1 inhibitor A922500 (HY-10038) was purchased from MCE; oleic acid (HY-N1446) was purchased from MCE; CPT1A inhibitors Etomoxir (HY-50202) and CP640186 (HY-15259) were purchased from MCE; and rabbit polyclonal antibody against FMDV VP1 protein (type O) (bs-41049R) was purchased from Beijing Biosynthesis Biotechnology Co., Ltd. (bioss).

Techniques: Infection

Journal: Redox Biology

Article Title: Activity-based CO 2 sensing using CarboSenR2 provides new insights into cellular metabolism

doi: 10.1016/j.redox.2026.104067

Figure Lengend Snippet: Assessment of CO 2 Production in CarboSenR2-Loaded C2C12 Myoblast Cells Following Exercise-Like Stimuli (A) Visual confirmation of myoblast-to-myotube differentiation following the stated differentiation protocol. Images are representative of three fields of view & three independent experiments (N = 3). (B) Quantitative plots showing the geometric mean of R-Dye fluorescence (575 ± 20 nm) in C2C12 myotube cells loaded with CarboSenR2, comparing cells stimulated with EPS to those maintained at rest for 1 h in a 5% CO 2 environment. CarboSenR2 was used at a concentration of 1.5 μM. Fluorescence was quantified using a Beckman Coulter CytoFlex LX in the PE channel (575 ± 20 nm). Data represent four independent experiments (N = 4) and are presented as fold change ± standard deviation relative to vehicle control. Statistical analysis was conducted using a paired two-tailed t -test. Statistical significance is indicated as follows: ∗∗p < 0.01. (C) Representative flow cytometry scatter plots showing the geometric mean of R-Dye fluorescence (575 ± 20 nm) in CarboSenR2-loaded C2C12 myoblast cells. (D) Quantitative plots showing the geometric mean of Biotracker 405 (440 ± 20 nm) in C2C12 myoblast cells comparing cells stimulated with EPS to those maintained at rest for 1 h in a 5% CO 2 environment. Biotracker 405 was used at a concentration of 100 nM. Fluorescence was quantified using a Beckman Coulter CytoFlex LX at the stated wave length. Data represent four independent experiments (N = 4) and are presented as fold change ± standard deviation relative to vehicle control. (E) Quantitative plots showing the geometric mean of R-Dye fluorescence (575 ± 20 nm) in C2C12 myoblast cells loaded with CarboSenR2, comparing cells stimulated with EPS to those maintained at rest for 1 h in a 5% CO 2 environment. CarboSenR2 was used at a concentration of 1.5 μM. Fluorescence was quantified using a Beckman Coulter CytoFlex LX in the PE channel (575 ± 20 nm). Data represent five independent experiments (N = 5) and are presented as fold change ± standard deviation relative to vehicle control. Statistical analysis was conducted using a paired two-tailed t -test. Statistical significance is indicated as follows: ∗∗∗p < 0.001. (F) Western blot analysis of phosphorylated Acetyl-CoA Carboxylase expression in cytosolic fractions of C2C12 myoblast cells following 1 h EPS stimulation. Data is expressed as fold change of protein expression normalised to total protein revert stain when comparing exercise to rest, and represents three independent experiments (N = 3). (G) Western blot analysis of nuclear PGC-1α expression in nuclear fractions of C2C12 myoblast cells following 1 h EPS stimulation. Data is expressed as fold change of protein expression normalised to total Lamin A/C content (nuclear loading control) when comparing exercise to rest, and represents three independent experiments (N = 3). Nuclear fractions were prepared as per methods. Lysates were probed using revert total protein stain and imaged in the 700 nm channel. Blots were incubated with a respective primary antibodies followed by a fluorescent secondary fluorophore-conjugated antibody and imaged in the 680-nm or 800-nm channel on an Li-COR imaging system. Relative intensities were quantified using Empiria Studio 3.0 Software. Image is representative of three independent experiments.

Article Snippet: Membranes were blocked in 5% milk in TBST for 1 h followed by overnight incubation in primary antibody (1:1000) at 4 °C (phospho Acetyl-CoA Carboxylase (Ser79) (CST-3661S, Cell Signalling), PGC-1a (A12348, ABclonal), Lamin A/C (CST-4777S, Cell Signalling), mitochondrial cytochrome c oxidase subunit 1 (ab14705, Abcam)).

Techniques: Fluorescence, Concentration Assay, Standard Deviation, Control, Two Tailed Test, Flow Cytometry, Western Blot, Expressing, Staining, Incubation, Imaging, Software