Journal: Tumour Virus Research

Article Title: Oncolytic virus hijacks GOT1 and pyrimidinosomes to fuel pyrimidine synthesis for replication in tumor cells

doi: 10.1016/j.tvr.2026.200342

Figure Lengend Snippet: | NDV infection promotes the assembly of pyrimidinosome involving GOT1 and pyrimidine synthases. (A) H1299 cells were co-transfected with GFP-GOT1 and mCherry-UMPS for 24 h, followed by mock infection or NDV infection (MOI = 0.1, 1, 5, 10, or UV-NDV for 12 hpi). Cells were then analyzed using confocal microscopy. (B) H1299 cells were co-transfected with GFP-GOT1 and mCherry-UMPS for 24 h, followed by mock infection or NDV infection (MOI = 1 for 6, 12, and 18 hpi). Cells were then analyzed using confocal microscopy. (C) Immunofluorescence analysis (IFA) of mock-infected and NDV-infected A549 cells (MOI = 1, 12 hpi), stained for endogenous GOT1, UMPS, and DHODH, alongside the mitochondrial marker protein Tom 20. (D-F) H1299 cells were co-transfected with GFP-GOT1 and mCherry-UMPS (D), GFP-DHODH and mCherry-UMPS (E), or GFP-GOT1 and mCherry-DHODH (F) for 24 h, followed by mock infection or NDV infection (MOI = 1, 12 hpi). Cells were then analyzed using confocal microscopy. (G-H) HEK293T cells were mock-infected or infected with NDV (MOI = 1) for 12 h. The cells were lysed and subjected to immunoprecipitation (IP) using GOT1 (G) and UMPS (H) antibodies, alongside with anti-IgG antibody as a control, followed by WB with anti-CAD, -UMPS, -DHODH and -GOT1 antibodies. (I-J) H1299 cells were co-transfected with Flag-CAD and HA-GOT1 for 24 h, followed by mock infection or NDV infection (MOI = 1, 12 hpi). The cells were lysed and subjected to IP using anti-Flag (I) or anti-HA (J) magnetic beads, followed by WB with anti-Flag and anti-HA antibodies. (K) H1299 cells were transfected with GFP-GOT1, mCherry-UMPS, or GFP-DHODH for 24 h, followed by NDV infection (MOI = 1, 12 hpi). FRAP analysis was then performed using confocal microscopy.

Article Snippet: Anti-Flag Magnetic Beads (Cat# HY-K0207), anti-HA Magnetic Beads (Cat# HY-K0201), Mycophenolate (Cat# HY-B0421), Leflunomide (Cat# HY-B0083), AG 2037 (Cat# HY-14530), Aminooxyacetic acid hemihydrochloride (Cat# HY-107994) were purchased from MedChemExpress (MCE).

Techniques: Infection, Transfection, Confocal Microscopy, Immunofluorescence, Staining, Marker, Immunoprecipitation, Control, Magnetic Beads

Journal: Tumour Virus Research

Article Title: Oncolytic virus hijacks GOT1 and pyrimidinosomes to fuel pyrimidine synthesis for replication in tumor cells

doi: 10.1016/j.tvr.2026.200342

Figure Lengend Snippet: | NDV infection promotes the assembly of pyrimidinosome involving GOT1 and pyrimidine synthases. (A) H1299 cells were co-transfected with GFP-GOT1 and mCherry-UMPS for 24 h, followed by mock infection or NDV infection (MOI = 0.1, 1, 5, 10, or UV-NDV for 12 hpi). Cells were then analyzed using confocal microscopy. (B) H1299 cells were co-transfected with GFP-GOT1 and mCherry-UMPS for 24 h, followed by mock infection or NDV infection (MOI = 1 for 6, 12, and 18 hpi). Cells were then analyzed using confocal microscopy. (C) Immunofluorescence analysis (IFA) of mock-infected and NDV-infected A549 cells (MOI = 1, 12 hpi), stained for endogenous GOT1, UMPS, and DHODH, alongside the mitochondrial marker protein Tom 20. (D-F) H1299 cells were co-transfected with GFP-GOT1 and mCherry-UMPS (D), GFP-DHODH and mCherry-UMPS (E), or GFP-GOT1 and mCherry-DHODH (F) for 24 h, followed by mock infection or NDV infection (MOI = 1, 12 hpi). Cells were then analyzed using confocal microscopy. (G-H) HEK293T cells were mock-infected or infected with NDV (MOI = 1) for 12 h. The cells were lysed and subjected to immunoprecipitation (IP) using GOT1 (G) and UMPS (H) antibodies, alongside with anti-IgG antibody as a control, followed by WB with anti-CAD, -UMPS, -DHODH and -GOT1 antibodies. (I-J) H1299 cells were co-transfected with Flag-CAD and HA-GOT1 for 24 h, followed by mock infection or NDV infection (MOI = 1, 12 hpi). The cells were lysed and subjected to IP using anti-Flag (I) or anti-HA (J) magnetic beads, followed by WB with anti-Flag and anti-HA antibodies. (K) H1299 cells were transfected with GFP-GOT1, mCherry-UMPS, or GFP-DHODH for 24 h, followed by NDV infection (MOI = 1, 12 hpi). FRAP analysis was then performed using confocal microscopy.

Article Snippet: Anti-Flag Magnetic Beads (Cat# HY-K0207), anti-HA Magnetic Beads (Cat# HY-K0201), Mycophenolate (Cat# HY-B0421), Leflunomide (Cat# HY-B0083), AG 2037 (Cat# HY-14530), Aminooxyacetic acid hemihydrochloride (Cat# HY-107994) were purchased from MedChemExpress (MCE).

Techniques: Infection, Transfection, Confocal Microscopy, Immunofluorescence, Staining, Marker, Immunoprecipitation, Control, Magnetic Beads

Journal: aBIOTECH

Article Title: The oxidation of ABI4 by RBOHD-derived reactive oxygen species integrates redox signaling into abscisic-acid and drought-stress responses

doi: 10.1016/j.abiote.2026.100037

Figure Lengend Snippet: RBOHD-produced H 2 O 2 mediates ABI4 sulfenylation. A Effect of H 2 O 2 concentration on ABI4 sulfenylation. Protoplasts from abi4 mutants transiently expressing ABI4-GFP were treated with the indicated concentrations of H 2 O 2 for 60 min. ABI4-GFP was immunoprecipitated using anti-GFP magnetic beads, and sulfenylation (SOH) levels were detected using an anti-SOH antibody. B Time-dependent sulfenylation of ABI4 following treatment with 10 μM H 2 O 2 . C-D ABA-induced sulfenylation of ABI4 in protoplasts from abi4 (C) and abi4 rbohd (D) mutants transiently expressing ABI4-GFP following treatment with 10 μM ABA for the indicated times. E Identification of the sulfenylated cysteine residue in ABI4. Protoplasts transiently expressing ABI4-GFP or cysteine-mutated variants were treated with H 2 O 2 , followed by immunoprecipitation and SOH detection as described above. Sulfenylation levels were normalized to the untreated control (set to 1.0). Data represent means ± SE from at least three independent biological replicates. Different letters indicate statistically significant differences ( P < 0.05, Duncan's multiple range test).

Article Snippet: These samples purified by with GFP magnetic beads (Smart-Lifesciences) were run on an SDS-PAGE gel, and the protein was transferred onto a polyvinylidene difluoride membrane.

Techniques: Produced, Concentration Assay, Expressing, Immunoprecipitation, Magnetic Beads, Residue, Control