Journal: Journal of Virology

Article Title: STING agonist diABZI confers protection against swine acute diarrhea syndrome coronavirus in neonatal mice by activating antiviral immunity

doi: 10.1128/jvi.01703-25

Figure Lengend Snippet: SADS-CoV infection has high lethality in C57BL/6J mice. Three-day-old, 7-day-old, and 3-week-old mice were infected with SADS-CoV, and 3-day-old infected mice were used as a control group ( n = 6). ( A ) Newborn C57BL/6J mice were inoculated with SADS-CoV via intraperitoneal injection. ( B ) Daily monitoring of survival rate. ( C ) Clinical scores were monitored daily, with clinical scoring based on . ( D ) Viral RNA in tissues collected from infected mice at 3 and 7 dpi was quantified using RT-qPCR. Data (mean ± SD) are from three independent triplicate experiments. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: WT C57BL/6J mice of different ages and C57BL/6J Smoc-STING KO mice were procured from Shanghai Model Organisms Center, Inc.

Techniques: Infection, Control, Injection, Quantitative RT-PCR

Journal: Journal of Virology

Article Title: STING agonist diABZI confers protection against swine acute diarrhea syndrome coronavirus in neonatal mice by activating antiviral immunity

doi: 10.1128/jvi.01703-25

Figure Lengend Snippet: The antiviral activity of diABZI against SADS-CoV is STING dependent. SADS-CoV or mock infection of neonatal C57BL/6J wild-type (WT) and C57BL/6J Smoc-STING KO (STING KO ) 3-day-old neonatal mice ( n = 6) was performed, followed by intraperitoneal injection (12 h post-viral infection) of corn oil containing 2.5 mg/kg diABZI (an agonist of the STING pathway) or DMSO dissolved in corn oil once per day for 3 days. The mice were euthanized after 3 days, and spleen tissues were collected for real-time PCR analysis, with a focus on the expression levels of the cytokines IFN-β ( A ), IFN-λ3 ( B ), IFIT1 ( C ), MX1 ( D ), IL-1β ( E ), and IFN-γ ( F ). Scale bar: 100 μm. Data (mean ± SD) are from three independent triplicate experiments. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. ( G ) Western blotting was used to evaluate STING and SADS-CoV N protein levels and the phosphorylation status of the STING protein in the spleen. ( H ) Immunofluorescence was used to detect the expression of STING and SADS-CoV N protein in the spleen. ( I ) Mouse mortality rates were recorded daily. ( J ) The expression of SADS-CoV genome copies in spleen tissues was assessed using RT-qPCR. Scale bar: 100 μm. Data (mean ± SD) are from three independent triplicate experiments. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. ( K ) Pathological examinations of the brain and spleen were performed via H&E staining. Scale bar: 100 μm.

Article Snippet: WT C57BL/6J mice of different ages and C57BL/6J Smoc-STING KO mice were procured from Shanghai Model Organisms Center, Inc.

Techniques: Activity Assay, Infection, Injection, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Phospho-proteomics, Immunofluorescence, Quantitative RT-PCR, Staining

Journal: Journal of Virology

Article Title: STING agonist diABZI confers protection against swine acute diarrhea syndrome coronavirus in neonatal mice by activating antiviral immunity

doi: 10.1128/jvi.01703-25

Figure Lengend Snippet: The diABZI protects mice from SADS-CoV-induced damage by promoting IFN production. Neonatal C57BL/6J wild-type (WT) mice were intraperitoneally administered IgG1 + diABZI ( n = 6) or anti-IFNAR (1 mg) + diABZI ( n = 6). Spleen tissues were collected for qPCR analysis to assess the mRNA expression levels of the interferon-stimulated genes MX1 ( A ), CXCL10 ( B ), and ISG15 ( C ). Data (mean ± SD) are from three independent triplicate experiments. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001. Additional neonatal WT mice received the same treatments followed by infection or mock infection with SADS-CoV. Immunohistochemistry was performed to detect SADS-CoV nucleocapsid (N) protein expression in tissues ( D ). Mouse body weight loss ( E ) and survival ( F ) were monitored daily. Scale bar: 100 μm.

Article Snippet: WT C57BL/6J mice of different ages and C57BL/6J Smoc-STING KO mice were procured from Shanghai Model Organisms Center, Inc.

Techniques: Expressing, Infection, Immunohistochemistry

Journal: Cell Death & Disease

Article Title: PRMT1-mediated methylation of UBE2m promoting calcium oxalate crystal-induced kidney injury by inhibiting fatty acid metabolism

doi: 10.1038/s41419-025-07888-3

Figure Lengend Snippet: A The violin plot showing the expression level of PRMT1 in various cell clusters from the NC and the KS groups. B , C The volcano plot of different expression genes or proteins assayed using RNA- or proteome sequencing of kidney tissues. PRMT1 was significantly increased at both the RNA and protein levels. D Representative immunohistochemical staining images (100× and 400×) and quantitative analysis ( F ) of the expression of PRMT1 in the nonfunctioning kidney of patients and in kidneys with CaOx crystals injury of mice. Scale bars = 50 μm. E Representative western blot banding and quantitative analysis ( G ) of the expression levels of PRMT1 and MMA in the NC and the Gly groups. H Representative images of immunofluorescence co-staining of PRMT1 (red) and renal tubular markers LTL, PNA, and DBA (green) in kidneys of mice. Scale bars = 50 μm. I Oil Red O staining of kidney sections from the indicated groups. J GO enrichment analysis of RNA-sequencing data comparing the expression levels between NC and the Gly group. K Heat map showing differential content of acyl-CoAs between the NC and the Gly groups. L GSEA enrichment analysis of differential proteins between the Gly group and the CKO group, assayed by proteome sequencing of kidney tissues. M Relative mRNA levels of key molecules of lipogenesis, fatty acid transport, and oxidation from the indicated groups. Significance was assessed using two-way ANOVA or t-tests. Data are presented as mean ± SD.

Article Snippet: PRMT1 floxed mice (C57BL/6J background, PRMT1fl/fl) were obtained from Shanghai Model Organisms.

Techniques: Expressing, Sequencing, Immunohistochemical staining, Staining, Western Blot, Immunofluorescence, RNA Sequencing

Journal: Cell Death & Disease

Article Title: PRMT1-mediated methylation of UBE2m promoting calcium oxalate crystal-induced kidney injury by inhibiting fatty acid metabolism

doi: 10.1038/s41419-025-07888-3

Figure Lengend Snippet: A Representative histology images of the indicated groups: Vonkossa staining indicates the area and location of calcium oxalate crystals deposition (400×); HE staining shows the degree of renal tubular injury (200×); TUNEL staining shows the relative number of apoptotic renal cells (200×). Scale bars = 50 μm. B , C Representative TEM images demonstrate the degree of mitochondrial damage and lipid droplet accumulation. Scale bars = 500 nm. D Representative western blot banding and quantitative analysis of the expression levels of ACOX1, CPT1a, CD36, and FATP2. E Mitochondrial oxidative capacity was measured in real time after knockdown of PRMT1 or COM treatment in HK-2 cells; Quantification of basal respiration, ATP production-coupled respiration, and maximal respiration. Significance was assessed using two-way ANOVA tests. Data are shown as mean ± SD.

Article Snippet: PRMT1 floxed mice (C57BL/6J background, PRMT1fl/fl) were obtained from Shanghai Model Organisms.

Techniques: Staining, TUNEL Assay, Western Blot, Expressing, Knockdown

Journal: Cell Death & Disease

Article Title: PRMT1-mediated methylation of UBE2m promoting calcium oxalate crystal-induced kidney injury by inhibiting fatty acid metabolism

doi: 10.1038/s41419-025-07888-3

Figure Lengend Snippet: A Representative histology images of the indicated groups: Vonkossa staining indicates the area and location of calcium oxalate crystal deposition (1X and 200X); HE staining shows the degree of renal tubular injury (200X); TUNEL staining shows the relative number of apoptotic renal cells (200X). Scale bars = 50 μm. B , C Representative TEM images demonstrate the degree of mitochondrial damage and lipid droplet accumulation. Scale bars = 500 nm. D Representative western blot bandings show the expression levels of ACOX1 and CPT1α. E Mitochondrial oxidative capacity was measured in real time after overexpression of PRMT1 or COM treatment in HK-2 cells; basal respiration, ATP production-coupled respiration, and maximal respiration were quantified. Significance was assessed using two-way ANOVA tests. Data are shown as mean ± SD.

Article Snippet: PRMT1 floxed mice (C57BL/6J background, PRMT1fl/fl) were obtained from Shanghai Model Organisms.

Techniques: Staining, TUNEL Assay, Western Blot, Expressing, Over Expression

Journal: Cell Death & Disease

Article Title: PRMT1-mediated methylation of UBE2m promoting calcium oxalate crystal-induced kidney injury by inhibiting fatty acid metabolism

doi: 10.1038/s41419-025-07888-3

Figure Lengend Snippet: A Silver-stained SDS-PAGE gel of the immunoprecipitation product. B MS/MS spectra of the peptide “FSPSGIFGAFQR-COOH.” Peaks in color are the detected b (green) and y (red) ions. C The volcano plot indicating potential target proteins of PRMT1. D Molecular docking pattern diagram of PRMT1 and UBE2m. E Representative images of immunofluorescence staining of UBE2m (green) and PRMT1 (red) in HK-2 cells treated with COM. Scale bars = 50 μm. F Reciprocal co-IP analysis of PRMT1 and UBE2m in HK-2 cells with COM treatment. IgG was used as a negative control. G Recombinant GST-PRMT1 was incubated with recombinant His-UBE2m, followed by GST-pulldown and immunoblotting analysis with GST and His antibodies. H Endogenous UBE2m was immunoprecipitated in cells from the indicated groups. The mono-methylation (me1) level of UBE2m was determined using immunoblotting. I PLA detection of PRMT1 and UBE2m interaction in HK-2 cells from the indicated groups. Scale bars = 50 μm. J The me1 level of UBE2m was determined using immunoblotting after UBE2m was immunoprecipitated in HK-2 cells with PRMT1 knockdown or overexpression.

Article Snippet: PRMT1 floxed mice (C57BL/6J background, PRMT1fl/fl) were obtained from Shanghai Model Organisms.

Techniques: Staining, SDS Page, Immunoprecipitation, Tandem Mass Spectroscopy, Immunofluorescence, Co-Immunoprecipitation Assay, Negative Control, Recombinant, Incubation, Western Blot, Methylation, Knockdown, Over Expression

Journal: Cell Death & Disease

Article Title: PRMT1-mediated methylation of UBE2m promoting calcium oxalate crystal-induced kidney injury by inhibiting fatty acid metabolism

doi: 10.1038/s41419-025-07888-3

Figure Lengend Snippet: A The scores of potential methylation sites of UBE2m predicted using the GPS-MSP tool. B MS identification of R169 mono-methylation of HA-UBE2m immunopurified by HA beads. C Sequence alignment of UBE2m protein from indicated species. D HA-UBE2m WT or different point mutants were co-transfected with FLAG-PRMT in HEK293T cells. The me1 level of UBE2m was measured by immunoblotting following immunopurification. E The me1 of immunopurified HA-UBE2m was measured using a site-specific antibody against R169 mono-methylation (meUBE2m (R169me1)). Antibody efficacy and specificity were examined by pre-incubating with the R169me1 peptide or the unmodified peptide prior to application. F Dot blot analysis of different amounts of R169me1 peptide or unmodified peptide by a site-specific antibody against meUBE2m (R169me1). G Endogenous UBE2m was immunoprecipitated in cells from the indicated groups. The R169me1 level of UBE2m was determined using immunoblotting. H The R169me1 level of UBE2m was determined using immunoblotting after UBE2m was immunoprecipitated in HK-2 cells with PRMT1 knockdown or overexpression. I Recombinant GST-PRMT1 and His-UBE2m proteins were incubated with or without SAM to detect the methylation of UBE2m mediated by PRMT1 in vitro. The reaction mixture was then subjected to CBB staining and immunoblotting with meUBE2m (R169me1) antibody. J Mitochondrial oxidative capacity was measured in real time after reintroduced UBE2m WT or RK treatment in UBE2m KO HK-2 cells; basal respiration, ATP production-coupled respiration, and maximal respiration were quantified. Significance was assessed using two-way ANOVA tests. Data are shown as mean ± SD.

Article Snippet: PRMT1 floxed mice (C57BL/6J background, PRMT1fl/fl) were obtained from Shanghai Model Organisms.

Techniques: Methylation, Sequencing, Transfection, Western Blot, Immu-Puri, Dot Blot, Immunoprecipitation, Knockdown, Over Expression, Recombinant, Incubation, In Vitro, Staining

Journal: Cell Death & Disease

Article Title: PRMT1-mediated methylation of UBE2m promoting calcium oxalate crystal-induced kidney injury by inhibiting fatty acid metabolism

doi: 10.1038/s41419-025-07888-3

Figure Lengend Snippet: A RT-PCR was used to detect the RNA levels of PPARγ from the indicated groups. Representative western blot banding and quantitative analysis show the expression levels of PPARγ from the indicated groups in mice ( B ) or in HK-2 cells ( C ). D Detection of endogenous ubiquitination levels of PPARγ in HK-2 cells with COM-treatment. E Detection of endogenous ubiquitination levels of PPARγ in CaOx crystals kidney injury in mice. F , G Detection of exogenous ubiquitination levels of PPARγ in HEK193T cells with PRMT1 knockdown or overexpression. H Detection of exogenous ubiquitination levels of PPARγ in HEK193T cells from the indicated groups. Representative western blot banding and quantitative analysis of the expression levels of NEDD4 from the indicated groups in mice ( I ) or HK-2 cells ( J ). K HK-2 cells were treated with MG132. Representative western blot banding and quantitative analysis showed the expression levels of PPARγ from the indicated groups. L PLA detection of NEDD4 and PPARγ interaction in HK-2 cells from the indicated groups. Scale bars = 50 μm. M Detection of exogenous ubiquitination levels of PPARγ in HEK193T cells with knockdown of NEDD4. N Detection of exogenous ubiquitination levels of PPARγ in HEK193T cells from the indicated groups. Significance was assessed using two-way ANOVA tests. Data are shown as mean ± SD.

Article Snippet: PRMT1 floxed mice (C57BL/6J background, PRMT1fl/fl) were obtained from Shanghai Model Organisms.

Techniques: Reverse Transcription Polymerase Chain Reaction, Western Blot, Expressing, Ubiquitin Proteomics, Knockdown, Over Expression

Journal: Cell Death & Disease

Article Title: PRMT1-mediated methylation of UBE2m promoting calcium oxalate crystal-induced kidney injury by inhibiting fatty acid metabolism

doi: 10.1038/s41419-025-07888-3

Figure Lengend Snippet: A Representative immunohistochemical staining images (400X) and quantitative analysis of the expression of M-UBE2m in the nonfunctioning kidney of patients with stones or the control. Scale bars = 50 μm. B Pearson correlation analysis of the expression of PRMT1 and M-UBE2m. C Representative immunohistochemical staining images (400X) and quantitative analysis of the expression of KIM-1 in the nonfunctioning kidney of patients with stones or the control. Scale bars = 50 μm. D – G Pearson correlation analysis of the expression of PRMT1 and eGFR ( D ), PRMT1 and KIM-1 ( E ), M-UBE2m and eGFR ( F ), M-UBE2m and KIM-1 ( G ). Significance was assessed using t-tests. Data are presented as mean ± SD.

Article Snippet: PRMT1 floxed mice (C57BL/6J background, PRMT1fl/fl) were obtained from Shanghai Model Organisms.

Techniques: Immunohistochemical staining, Staining, Expressing, Control

Journal: Translational Psychiatry

Article Title: CCK2R regulates METH-induced CPP acquisition within VTA-BLA-BNST circuit in male mice

doi: 10.1038/s41398-026-03982-y

Figure Lengend Snippet: (A) Schematic of METH CPP procedure. (B) The expression of CCK1R mRNA. n = 5 mice. (C) The expression of CCK2R mRNA. BLA: Unpaired t test, t = 4.180, P = 0.0031, n = 5 mice. (D) The expression of CCKR mRNA in BLA after METH training. n = 5 mice. (E) Top: Representative immunoblots of CCK1R. Bottom: Relative amounts of CCK1R quantified by densitometry, in BLA. Unpaired t test, t = 0.317, P = 0.759, n = 5 mice. (F) Top: Representative immunoblots of CCK2R. Bottom: Relative amounts of CCK2R quantified by densitometry, in BLA. Unpaired t test, t = 2.620, P = 0.031, n = 5 mice. (G) Schematic of METH CPP procedure. (H) Schematic diagram of virus injection in BLA. Scale bar=200μm . (I, J) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (3,44) = 21.94, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (K, L) In vitro electrophysiology: AP spiking patterns evoked by 400 pA current injections and spike numbers and thresholds for AP firing. Two-way RM ANOVA ANOVA, F treatment (3,44) = 31.20, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (M, N) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (3,44) = 12.32, P <0.001; Tukey’s post hoc comparison, Vector+Saline vs Vector+METH: P <0.001, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. All data are means ± SEM, * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: CCK flox/flox (C57BL/6 J) and Vglut-Cre (C57BL/6 J) mice were obtained from the Shanghai Model Organisms Center, while CCK1R flox/flox (C57BL/6 J) and CCK2R flox/flox (C57BL/6 J) genotypes were procured from the Nanjing Biological Research Institute.

Techniques: Expressing, Western Blot, Virus, Injection, Comparison, Plasmid Preparation, In Vitro, Saline, In Vivo

Journal: Translational Psychiatry

Article Title: CCK2R regulates METH-induced CPP acquisition within VTA-BLA-BNST circuit in male mice

doi: 10.1038/s41398-026-03982-y

Figure Lengend Snippet: (A) Schematic of METH CPP procedure. (B) Schematic diagram of virus injection in VTA and BLA. Scale bar=200μm . (C, D) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (2,33) = 21.46, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (E, F) Representative traces of NMDAR and AMPAR mediated currents, and the bar chart of AMPAR/NMDAR ratio in BLA. Ordinary one-way ANOVA, F treatment (2,33) = 23.42, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (G, H) Representative traces of paired pulse and the bar chart of PPR in BLA. Ordinary one-way ANOVA, F treatment (2,33) = 11.96, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (I, J) The averaged power spectrograms of LFP and bar chart of theta oscillation(3-8 Hz) in BLA. Ordinary one-way ANOVA, F treatment (2,33) = 44.18, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. (K) Representative images of dendritic spines. (L) Bar chart of dendritic spine density in BLA. Ordinary one-way ANOVA, F treatment (2,15) = 12.15, P <0.001, n = 6 cells from 3 mice. (M) Density bar chart of thin, mushroom and stubby dendritic spine in BLA. Ordinary one-way ANOVA: thin dendritic spine, F treatment (2,15) = 17.43, P <0.001; mushroom dendritic spine, F treatment (2,15) = 6.915, P = 0.0074. All data are means ± SEM. * P < 0.5; ** P < 0.01; *** P < 0.001.

Article Snippet: CCK flox/flox (C57BL/6 J) and Vglut-Cre (C57BL/6 J) mice were obtained from the Shanghai Model Organisms Center, while CCK1R flox/flox (C57BL/6 J) and CCK2R flox/flox (C57BL/6 J) genotypes were procured from the Nanjing Biological Research Institute.

Techniques: Virus, Injection, Comparison, Plasmid Preparation

Journal: Translational Psychiatry

Article Title: CCK2R regulates METH-induced CPP acquisition within VTA-BLA-BNST circuit in male mice

doi: 10.1038/s41398-026-03982-y

Figure Lengend Snippet: (A) Schematic of METH CPP procedure. (B) Schematic diagram of virus injection in VTA, BLA and BNST. Scale bar=200μm . (C, D) Heat map and score bar chart of METH CPP. Two-way RM ANOVA, F treatment (2,27) = 16.33, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 mice. (E, F) In vitro electrophysiology: AP spiking patterns and spike numbers and thresholds for AP firing in BNST. Two-way RM ANOVA, F treatment (2,33) = 10.29, P <0.001, Tukey’s post hoc comparison, hM4Di+METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (G, H) In vivo electrophysiology: AP spiking patterns and bar chart of AP frequency. Ordinary one-way ANOVA, F treatment (2,33) = 7.431, P = 0.0022, Tukey’s post hoc comparison, hM4Di+METH vs Vector+METH: P = 0.0418, n = 12 cells from 3 mice. (I, J) Representative traces of NMDAR and AMPAR mediated currents, and the bar chart of AMPAR/NMDAR ratio in BNST. Ordinary one-way ANOVA, F treatment (2,33) = 26.06, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 4 mice. (K, L) Representative traces of paired pulse and the bar chart of PPR in BNST. Ordinary one-way ANOVA, F treatment (2,33) = 11.49, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P = 0.0047, n = 12 cells from 4 mice. (M, N) The averaged power spectrograms of LFP and bar chart of theta oscillation(3-8 Hz) in BNST. Ordinary one-way ANOVA, F treatment (2,33) = 24.14, P <0.001; Tukey’s post hoc comparison, CCK2R-KO + METH vs Vector+METH: P <0.001, n = 12 cells from 3 mice. (O) Representative images of dendritic spines. (P) Bar chart of dendritic spine density in BNST. Ordinary one-way ANOVA, F treatment (2,15) = 8.297, P = 0.0037, n = 6 cells from 3 mice. (Q) Density bar chart of thin, mushroom and stubby dendritic spine in BNST. Ordinary one-way ANOVA: thin dendritic spine, F treatment (2,15) = 9.296, P = 0.0024; mushroom dendritic spine, F treatment (2,15) = 7.405, P = 0.0058. All data are means ± SEM. * P < 0.5; ** P < 0.01; *** P < 0.001.

Article Snippet: CCK flox/flox (C57BL/6 J) and Vglut-Cre (C57BL/6 J) mice were obtained from the Shanghai Model Organisms Center, while CCK1R flox/flox (C57BL/6 J) and CCK2R flox/flox (C57BL/6 J) genotypes were procured from the Nanjing Biological Research Institute.

Techniques: Virus, Injection, Comparison, Plasmid Preparation, In Vitro, In Vivo