Journal: bioRxiv

Article Title: MOV10 inhibits SADS-CoV replication by enhancing TRIM24-mediated K63-linked TRAF3 ubiquitination and this inhibition is antagonized by viral N protein

doi: 10.64898/2026.01.09.698592

Figure Lengend Snippet: MOV10 promoted TRAF3 K63-linked polyubiquitination dependent on E3 ligase TRIM24. (A) HEK293T cells were co-transfected with Myc-hMOV10 and HA-TRIM24 for 24 h. Cell lysates were harvested and subjected to immunoprecipitation using HA Ab. (B) Co-localization of MOV10 and TRIM24. Plasmids expressing Myc-MOV10 (red) and HA-TRIM24 (green) were co-transfected into Vero E6 cells, which were then infected or not with SeV for 9 h. Merged images show co-localization of these proteins. Nuclei are highlighted by DAPI staining (blue). (Bar: 5 µm). (C) HEK293T cells were transfected with plasmids expressing Myc-hMOV10 or EV and infected or not with SeV. Protein expression of TRIM24, LaminA/C, and GAPDH in cytoplasmic (CE) and nuclear (NE) fractions was assessed by western blotting. The density of TRIM24 bands relative to GAPDH was calculated using grayscale analysis. (D) HEK293T cells were co-transfected with plasmids expressing HA-TRIM24, Flag-TRAF3, and Myc-hMOV10. Cells were harvested for Co-IP using HA Ab, with IgG Ab as the negative control. (E) Plasmids encoding HA-TRIM24, Flag-TRAF3, or Myc-hMOV10 (0, 1 or 3 μg) were co-transfected into HEK293T cells. Cell lysates were collected and then subjected to immunoprecipitation using HA Ab. (F) HEK293T cells were transfected with either siTRIM24 or siNC (50 nM) for 36 h, followed by co-transfection with Flag-TRAF3, HA-K63, or Myc-hMOV10. Lysates were subjected to immunoprecipitation using Flag Ab. (G and H) WT (TRIM24 +/+ ) and TRIM24 knockout (TRIM24 −/− ) HEK293T cells were co-transfected with Flag-TRAF3, HA-K63, or Myc-hMOV10. Lysates were subjected to immunoprecipitation using Flag Ab. (I and J) TRIM24 +/+ and TRIM24 −/− HEK293T cells were transfected with either siMOV10 or siNC, followed by co-transfection with Flag-TRAF3 and HA-K63. Lysates were subjected to immunoprecipitation using Flag Ab.

Article Snippet: Abs against MOV10 (10370-1-AP), TRAF3 (18099-1-AP), TRIM24 (14208-1-AP) and HA (51064-2-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Transfection, Immunoprecipitation, Expressing, Infection, Staining, Western Blot, Co-Immunoprecipitation Assay, Negative Control, Cotransfection, Knock-Out

Journal: bioRxiv

Article Title: MOV10 inhibits SADS-CoV replication by enhancing TRIM24-mediated K63-linked TRAF3 ubiquitination and this inhibition is antagonized by viral N protein

doi: 10.64898/2026.01.09.698592

Figure Lengend Snippet: SADS-CoV N protein degraded TRAF3 by catalyzing K48-linked polyubiquitination. (A) IPI-2I cells were infected with SADS-CoV (MOI = 0.1). Cells were collected at 6, 12, 24, and 36 h and detected by western blotting using Abs against TRAF3, MOV10, TRIM24, SADS-CoV N protein and GAPDH. (B) TRAF3 interacted with SADS-CoV N protein. HEK293T cells were co-transfected with Flag-TRAF3 and GFP-N. Cell extracts were subjected to immunoprecipitation using Flag Ab. (C) Co-localization of TRAF3 and SADS-CoV N protein. Plasmids expressing Flag-TRAF3 (red) and Myc-N (green) were transfected or co-transfected into Vero E6 cells. Merged images show co-localization of these proteins. Nuclei are highlighted by DAPI staining (blue). (Bar: 5 µm). (D) Plasmids encoding GFP-N, HA-pMOV10, and Flag-TRAF3 were transfected or co-transfected into HEK293T cells, respectively. Lysates were subjected to immunoprecipitation using HA Ab. (E) HEK293T cells were transfected with plasmids encoding Myc-N or EV. Lysates were subjected to immunoprecipitation using MOV10 Ab, with IgG Ab as the negative control. (F) IPI-2I cells were infected with SADS-CoV (MOI = 0.1) and treated with CQ, MG132, Z-VAD-FMK, or DMSO (all 20 μM) for 5 h before sample collection. Western blotting was used to detect expression of TRAF3, SADS-CoV N protein and GAPDH. (G) HEK293T cells were transfected or co-transfected with plasmids encoding Flag-TRAF3, HA-Ub, and Myc-N. Lysates were collected and then subjected to immunoprecipitation using Flag Ab. (H) HEK293T cells were co-transfected with plasmids encoding Flag-TRAF3, Myc-N, HA-K48 or HA-K63. Lysates were collected and then subjected to immunoprecipitation using Flag Ab. (I) HEK293T cells were co-transfected with Myc-N, HA-K48, Flag-TRAF3 or Flag-TRAF3 MATH. Lysates were subjected to immunoprecipitation using Flag Ab.

Article Snippet: Abs against MOV10 (10370-1-AP), TRAF3 (18099-1-AP), TRIM24 (14208-1-AP) and HA (51064-2-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Infection, Western Blot, Transfection, Immunoprecipitation, Expressing, Staining, Negative Control

Journal: bioRxiv

Article Title: MOV10 inhibits SADS-CoV replication by enhancing TRIM24-mediated K63-linked TRAF3 ubiquitination and this inhibition is antagonized by viral N protein

doi: 10.64898/2026.01.09.698592

Figure Lengend Snippet: Models of MOV10 mechanisms in counteracting SADS-CoV and viral N protein antagonizing host innate immunity. To inhibit SADS-CoV replication, the host factor MOV10 enhanced the catalysis of K63-linked polyubiquitination of TRAF3 by E3 ubiquitin ligase TRIM24, thereby activating the RIG-I-mediated antiviral signaling pathway. On the other hand, the SADS-CoV N protein suppressed RIG-I-mediated antiviral signaling by promoting K48-linked polyubiquitination, leading to degradation of TRAF3.

Article Snippet: Abs against MOV10 (10370-1-AP), TRAF3 (18099-1-AP), TRIM24 (14208-1-AP) and HA (51064-2-AP) were purchased from Proteintech (Wuhan, China).

Techniques: Ubiquitin Proteomics

Journal: Food & Function

Article Title: Punicalagin is the key pomegranate polyphenol inhibiting gut microbial trimethylamine (TMA) production from l -carnitine in an in vitro human colon model

doi: 10.1039/d5fo04781a

Figure Lengend Snippet: Effects of pomegranate polyphenols and gum Arabic at 2 mg mL −1 on in vitro l -carnitine metabolism. Average percentages of (A) l -carnitine, (B) γ-butyrobetaine (γ-BB), and (C) trimethylamine (TMA) relative to initial l -carnitine concentration are displayed over 48 hours. Results are shown as mean ± SD from 2–5 donors with 1–4 biological replicates each. Statistical analysis employed linear mixed models, including treatment as fixed effects with random intercepts for donors (* p < 0.05, ** p < 0.01, *** p < 0.001), to measure significant differences between the polyphenol treatments (high-throughput model) and the control. High-throughput and batch colon models were inoculated with 1% faecal inoculum from a healthy donor, 2 mM l -carnitine, and the treatment. After collection, samples were directly stored at −80 °C until LC-MS/MS quantification using isotope-labelled internal standards. The dashed line indicates the results obtained for treatments at a comparable dose of pomegranate extract (22.8 mg mL −1 ) and l -carnitine but in a pH-controlled in vitro batch colon model, as described in a previously published report.

Article Snippet: Individual pomegranate polyphenols were obtained from different suppliers: punicalagin was sourced from BOC Science (CAS 65995-63-3), punicalin from Apollo Scientific (CAS 65995-64-4), and Merck provided ellagic acid (CAS 476-66-4) and gallic acid (CAS 149-91-7).

Techniques: In Vitro, Concentration Assay, High Throughput Screening Assay, Control, Liquid Chromatography with Mass Spectroscopy

Journal: The Journal of Organic Chemistry

Article Title: 5(6)- anti -Substituted-2-azabicyclo[2.1.1]hexanes. A Nucleophilic Displacement Route

doi: 10.1021/jo901725k

Figure Lengend Snippet: Nucleophilic Substitutions of Bromide 10.

Article Snippet: To show that the benzyl group could be removed without destruction of the strained ring, alcohol 24 was hydrogenolyzed and the resulting amine was protected by reaction with (BOC) 2 O to give N -BOC alcohol 25 (92%). fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window SCHEME 3 caption a7 Reactions of Monobromides 10. table ft1 table-wrap mode="anchored" t5 caption a7 Open in a separate window entry substrate reagent conditions product X yield (%) 1 10 AgOAc DMF/60 °C/12h 24a OAc 54 2 10 AgF DMF/70 °C/24h 19 - 42 a 3 10 NaF DMF/70 °C/12h 10 Br 88 4 10 NaN 3 DMF/70 °C/12h 27 N 3 51 5 10 LiNu b DMF/70 °C/8d 29 Nu b 55 6 10 NaI c DMF/70 °C/3d 31 I 50 d 7 10 CsOAc e DMSO/70 °C/6h 24a OAc 90 8 10 NaF DMSO/70 °C/12h 10 Br 82 9 24b BAST CH 2 Cl 2 /40 °C/12h 26 F 62 10 10 AgF CH 3 NO 2 /50 °C/12h 26 F 80 11 10 NaN 3 DMSO/70 °C/5h 27 N 3 88 12 10 NaSPh DMSO/60 °C/5h 30 SPh 77 13 10 NaI f Acetone/reflux/4d 31 I 74 g Open in a separate window a Bromide 10 was stable in DMF/70 °C/12h; 90% recovery, no 19 formed. b Nu = N -imidazole. c 3 Equiv. d Admixed with unreacted bromide 10 . e To separate samples of the monobromide 10 in DMSO-d6 there was added 1.5 equiv of either NaOAc (sample A) or CsOAc (sample B).

Techniques: