anti disc1 antibody  (Santa Cruz Biotechnology)

Journal: Neural Regeneration Research

Article Title: Mitochondrial dynamics dysfunction and neurodevelopmental disorders: From pathological mechanisms to clinical translation

doi: 10.4103/NRR.NRR-D-24-01422

Figure Lengend Snippet: Mitochondrial axonal transport and plasticity abnormalities in neurodevelopmental disorders. In neurons, mitochondria travel long distances along microtubules driven by Kinesin-1 and the reverse dynein. Milton, also known as TRAK, acts as a linker between Miro, a protein anchored to the outer mitochondrial membrane, and the motor. (A) A high concentration of Ca 2+ activates Miro recruitment to the anchoring protein Synaphili, thereby preventing mitochondrial motility. DISC1 can bind to Synaphili and relieve the restriction on mitochondrial movement. When DISC1 is abnormal, it leads to schizophrenia and bipolar disorder. (B) Abnormalities in connexin TRAK can induce seizures and fatal encephalopathy. Created with BioRender.com. AD: Alzheimer’s disease; ALS: amyotrophic lateral sclerosis; DISC1: disrupted in schizophrenia 1; Dynein: a type of motor protein; Kinesin: a type of motor protein; Miro: mitochondrial Rho GTPase 1; SNPH: synphilin-1; Syntaphilin: a protein that interacts with synphilin-1; TRAK: trak protein family.

Article Snippet: Genetic analysis showed that the DISC1 gene is considered to be a highly associated genetic factor in schizophrenia in humans (Rittenhouse et al., 2021).

Techniques: Membrane, Concentration Assay

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Venn diagram showing the overlap of differentially expressed genes (DEGs) identified through transcriptomic and proteomic analyses of ZIKV-infected HTR8 cells compared to non-infected controls. The overlapping area indicates shared DEGs between the two datasets. b − d HTR8 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 hours ( h ) and then infected with ZIKV at a multiplicity of infection (MOI) of 1. Viral mRNA levels, protein expression and titers were assessed on day 1 (D1) and day 2 (D2) post-infection using qRT-PCR ( b ), Western blot ( c ), and plaque assay ( d ). e − g U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h and infected with ZIKV at a MOI of 1. Viral mRNA levels, protein expression and titers were measured on D1 and D2 post-infection using qRT-PCR ( e ), Western blot ( f ), and plaque assay ( g ). h , i HTR8 cells ( h ) or U251 cells ( i ) were transfected with siRNA targeting DISC1 for 24 h prior to ZIKV infection at a MOI of 1. Viral protein was analyzed on D1 and D2 post-infection using Western blot. j − m HTR8 cells ( j , k ) or U251 cells ( l , m ) were transfected with siRNA targeting DISC1 for 24 h prior to ZIKV infection at a MOI of 1. Viral mRNA levels and viral loads were analyzed on D1 and D2 post-infection using qRT-PCR ( j , l ) and plaque assay ( k , m ). Data shown in ( b , d – e , g , j – m ) are from one representative experiment out of three independent replicates with similar results, each including 3 biological replicates per group ( n = 3). Images in ( c − i ) are from one representative experiment out of three independent replicates with similar results ( n = 3). All the data were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using Student’s unpaired two-tailed t test. Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Infection, Transfection, Expressing, Quantitative RT-PCR, Western Blot, Plaque Assay, Two Tailed Test

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Schematic representation of the experiment set up. 6-8-week-old WT or Disc1 KD mice were treated with 2 mg MAR1-5A3 on the day prior to infection and then intraperitoneally (i.p.) inoculated with PBS or 1 × 10 6 PFU of ZIKV. Mouse tissues were collected on D2 and D6 post-infection. b − h Viral titers in plasma ( b ) and uterus ( h ) were determined by plaque assay, and mRNA levels in blood cell ( c ), spleen ( d ), brain ( e ), testis ( f ) and uterus ( g ) were measured by qRT-PCR on D2 and D6 post-infection. i , j Primary cells isolated from WT or Disc1 KD mice were induced to differentiate into macrophages using mouse macrophage colony-stimulating factor. Cells were pre-incubated with 20 μg/mL anti-IFNAR1 antibody MAR1-5A3 for 6 h, followed by infection with ZIKV at a MOI of 1. Viral mRNA levels and titers were measured on D1 and D3 post-infection using qRT-PCR ( i ) and plaque assay ( j ). Data for plasma ( b ), blood cells ( c ), spleen ( d ), and brain ( e ) are one representative experiment out of two independent replicates with similar results, each including 6 mice per group ( n = 6), consisting of 3 males and 3 females. Data for the testis ( f ) are one representative experiment out of two independent replicates with similar results, each including 6 male mice per group ( n = 6). Data for the uterus ( g , h ) are one representative experiment out of two independent replicates with similar results, each including 6 female mice per group ( n = 6). Data shown in ( i , j ) are one representative experiment out of two independent replicates with similar results, each including 4 biological replicates per group ( n = 4). Data shown in ( b , e , j ) did not follow a normal distribution according to the Shapiro-Wilk test, and statistical analysis was performed using two-tailed Mann-Whitney test. Data shown in ( c – d , f – i ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using Student’s unpaired two-tailed t test. Data are presented as means ± SD. Source data are provided as a Source Data file. Figure 2a was created in BioRender. Hl, Z. (2025) https://BioRender.com/bh2c5ad .

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Infection, Clinical Proteomics, Plaque Assay, Quantitative RT-PCR, Isolation, Incubation, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Schematic representation of the experimental setup. One day prior to infection, 2 mg of the anti-IFNAR1 antibody MAR1-5A3 was administered to 8-10-week-old WT dams and Disc1 KD dams on E5.5. On E6.5, dams were i.p. inoculated with either PBS or 1 × 10 7 PFU of ZIKV. Placenta and fetal head were harvested on E13.5. b Representative images of E13.5 uteri (upper panel) and fetuses (lower panel). Partial demise and growth restriction was shown in ZIKV-infected WT and Disc1 KD pregnant dams. Red arrows indicate the placental residues, and green arrows show the growth restriction of fetuses. c Resorption rates were analyzed on E13.5. d− g The weight ( d ), CRL ( e ), OFD ( f ) and size ( g , CRL × OFD) of fetuses were measured on E13.5. h , i ZIKV mRNA levels in placentas ( h ) and fetal heads ( i ) of WT and Disc1 KD mice was measured by qRT-PCR. j ZIKV mRNA levels in the spleen of in WT or Disc1 KD dams were measured by qRT-PCR. Data shown in ( d − i ) are one representative experiment out of two independent replicates with similar results, each including 4 pregnant dams per group ( n = 4). The offspring analyzed were NF-WT ( n = 36), NF-KD ( n = 37), ZIKV-WT ( n = 28), and ZIKV-KD ( n = 20). Data shown in j are one representative experiment out of two independent replicates with similar results, each including 4 pregnant dams per group ( n = 4). Data shown in ( d − i ) did not follow a normal distribution according to the Shapiro-Wilk test, and statistical analysis was performed using two-tailed Kruskal-Wallis test followed by Dunn’s multiple comparison test ( d − g ) or two-tailed Mann-Whitney test ( h , i ). Data shown in e and j were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test ( e ) or Student’s unpaired two-tailed t test ( j ). Data are presented as means ± SD. Source data are provided as a Source Data file. Figure 3a was created in BioRender. Hl, Z. (2025) https://BioRender.com/dvzm7vs .

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Infection, Quantitative RT-PCR, Two Tailed Test, Comparison, MANN-WHITNEY

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Schematic representation of the experiment set up. On the second day after birth, WT and Disc1 KD neonatal mice were intracranially injected with 200 PFU of ZIKV or an equal volume of PBS. Brains were collected on D3, D6 and D9 post-infection, as well as at 6 weeks of age. b ZIKV mRNA levels in WT and Disc1 KD brains were assessed by qRT-PCR on D3, D6 and D9 post-infection, as well as at 6 weeks of age. c , d Body weight ( c ) and brain weight ( d ) of WT and Disc1 KD mice were measured on D3, D6, D9 post-infection, as well as at 6 weeks of age. e , f Representative images of WT and Disc1 KD mice brains on D3, D6, D9 post-infection ( e ), as well as at 6 weeks of age ( f ). The yellow scale bar indicates consistent size within the same age group. g The cerebral size of WT and Disc1 KD mice was measured on D3, D6, D9 post-infection, as well as at 6 weeks of age. Data shown in ( b − d , g ) are collected from two independent replicates, including 8 mice per group ( n = 8), consisting of 4 males and 4 females. Images in ( e , f ) are one representative experiment out of two independent replicates with similar results. Data shown in ( b ) did not follow a normal distribution according to the Shapiro-Wilk test, and statistical analysis was performed using two-tailed Mann-Whitney test. Data shown in ( c – d , g ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test. Data are presented as means ± SD. Source data are provided as a Source Data file. Figure 4a was created in BioRender. Hl, Z. (2025) https://BioRender.com/s3be69y .

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Injection, Infection, Quantitative RT-PCR, Two Tailed Test, MANN-WHITNEY, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a On the second day after birth, WT and Disc1 KD neonatal mice were intracranially injected with 200 PFU of ZIKV or an equal volume of PBS, and behavioral tests were conducted when the mice reached 6 weeks of age. The spontaneous alternation rate of WT and Disc1 KD mice in the Y-maze test. b The passing time of WT and Disc1 KD mice in the balance beam test. c , d Proportion of entries into the open arms ( c ) and the percentage of time spent in the open arms ( d ) by WT and Disc1 KD mice in the elevated plus maze test. e , f The time of WT and Disc1 KD mice spent on socializing with Stranger 1 ( e ) and Stranger 2 ( f ) during the 3-chamber test. Data shown in ( a − d ) are collected from one independent experiment, each including 8 mice per group ( n = 8), consisting of 4 males and 4 females. Data shown in ( e , f ) are collected from one independent experiment, each including 10 mice per group ( n = 10), consisting of 5 males and 5 females. Data shown in ( a - b , d – f ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test. Data shown in c did not follow a normal distribution according to the Shapiro-Wilk test, statistical analysis was performed using two-tailed Kruskal-Wallis test followed by Dunn’s multiple comparison test. Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Injection, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a KEGG pathway enrichment of DEGs in ZIKV-infected U251 cells transfected with HA-DISC1 (1 μg) plasmid compared with the control group transfected with pcDNA. b KEGG pathway enrichment of differentially expressed proteins identified by IP/MS analysis in U251 cells overexpressing HA-DISC1 (5 μg) plasmid compared with the control group transfected with pcDNA. c Chord diagram illustrates the proteins identified in the IP/MS analysis. d U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h followed by ZIKV infection. The protein levels of AMPKα, pAMPKα, mTOR, pmTOR, P62 and LC3A/B were assessed by Western blot at 0, 12, 24, and 36 h post-infection. e U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h, followed by ZIKV infection for 6 h and treated with DMSO, rapamycin (25 μM), and MHY1485 (10 μM) for 24 h. ZIKV E expression was assessed by Western blot. f U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h, followed by ZIKV infection for 6 h and treated with DMSO, autophagosome inhibitor 3-MA (1 mg/ml), and lysosome inhibitor CQ (100 μM) for 24 h. ZIKV E expression was assessed by Western blot. KEGG pathway enrichment analyses in ( a , b ) were performed using a one-tailed Fisher’s exact test. Images in ( d − f ) are one representative experiment out of three independent replicates with similar results ( n = 3). Source data are provided as a Source Data file.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Infection, Transfection, Plasmid Preparation, Control, Protein-Protein interactions, Western Blot, Expressing, One-tailed Test

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Conserved LIR motifs (amino acids 210 FSFI 213) in DISC1 were highlighted in yellow, which were mutated into AAAA in DISC1 mutant. b 293T cells were co-transfected with HA-DISC1 (5 μg) or HA-DISC1 mutant (5 μg) and GFP-LC3 (5 μg) plasmids for 48 h. Cellular lysates were subjected to immunoprecipitation with anti-Flag or anti-HA magnetic beads and Western blot assays using the indicated antibodies. c − e U251 cells were transfected with HA-DISC1 (1 μg) or HA-DISC1 mutant (1 μg) plasmids for 24 h, followed by ZIKV infection for 24 and 48 h. Viral mRNA levels, protein expression and titers were measured on D1 and D2 post-infection by qRT-PCR ( c ), Western blot ( d ), and plaque assays ( e ). Images in ( b , d ) are from one representative experiment out of three independent replicates with similar results ( n = 3). Data shown in ( c , e ) are from one representative experiment out of three independent replicates with similar results, each including 3 biological replicates per group ( n = 3). Data shown in ( c , e ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test. Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Mutagenesis, Transfection, Immunoprecipitation, Magnetic Beads, Western Blot, Infection, Expressing, Quantitative RT-PCR, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Primary cortical neurons were isolated from neonatal mice (P0–P1) and induced to differentiate in vitro. On day 7 of differentiation, neurons exhibiting neurite outgrowth were observed under bright-field microscopy. Confocal imaging showed neuronal nuclei stained with DAPI (blue), and endogenous Neun labeled with an anti-Neun antibody (red). Scale bar = 50 μm. b , c WT and Disc1 KD primary cortical neurons were pre-incubated with 20 μg/mL MAR1-5A3 for 6 h, followed by infection with ZIKV at a MOI of 1. Viral mRNA levels and titers were measured on D1 and D2 post-infection using qRT-PCR ( b ) and plaque assay ( c ). d WT and Disc1 KD primary cortical neurons were pre-incubated with 20 μg/mL MAR1-5A3 for 6 h, followed by infection with ZIKV at a MOI of 1. The protein levels of ZIKV E protein, DISC1, AMPKα, pAMPKα, mTOR, pmTOR, P62 and LC3A/B were assessed by Western blot on D1 and D2 post-infection. e, f The ratios of pAMPKα to AMPKα ( e ) and pmTOR to mTOR ( f ) were determined by comparing the corresponding protein band intensities at each time point in Fig. 8d. g − i Quantification of P62 ( g ), LC3A/BⅠ ( h ) and LC3A/BⅡ ( i ) protein levels relative to GAPDH at each time point in Fig. 8d. j , k Placentas ( j ) and fetal heads ( k ) from non-infected and ZIKV-infected WT and Disc1 KD mice were collected on E13.5. The protein levels of DISC1, AMPKα, pAMPKα, mTOR, pmTOR, P62 and LC3A/B were assessed by Western blot. l − o The ratios of pAMPKα to AMPKα ( l, m ) and pmTOR to mTOR ( n , o ) were determined by comparing the corresponding protein band intensities at each time point in Fig. 8j ( l, n ) and Fig. 8k ( m , o ). p − s Quantification of P62 ( p , q ) and LC3A/BⅡ ( r , s ) protein levels relative to GAPDH at each time point in Fig. 8j ( p , r ) and Fig. 8k ( q , s ). Data shown in ( b , c ) are from one representative experiment out of three independent replicates with similar results, each including 3 biological replicates per group ( n = 3). Images in ( a , d , j – k ) are from one representative experiment out of three independent replicates with similar results ( n = 3). Data shown in ( e − i , l − s) are collected from three independent experiments ( n = 3). Data shown in ( b − c , e − i , l − s ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using Student’s unpaired two-tailed t test ( b , c ) or two-tailed one-way ANOVA followed by Tukey’s multiple comparison test ( e − i , l − s ). Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Isolation, In Vitro, Microscopy, Imaging, Staining, Labeling, Incubation, Infection, Quantitative RT-PCR, Plaque Assay, Western Blot, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a 6-8-weeks-old WT or Disc1 KD mice were treated with 2 mg MAR1-5A3 on the day prior to infection and then intraperitoneally (i.p.) inoculated with PBS or 1 × 10 6 PFU of ZIKV. Mouse brains were collected on D6 post-infection. Representative H&E-stained sections of the hippocampal DG and CA3 regions are shown. Black arrows indicate shrunken, hyperchromatic neurons with indistinct boundaries Scale bar = 100 μm. b Representative immunofluorescence-stained sections of the hippocampal DG region. Cell nuclei were stained using DAPI (blue). Endogenous Neun was labeled with anti-Neun antibody (red). Endogenous LC3A/B was labeled with anti-LC3A/B antibody (yellow). Scale bar = 100 μm. Representative images in ( a , b ) are from one independent experiment, including 6 mice per group ( n = 6), consisting of 3 males and 3 females.

Article Snippet: The Disc1 knockdown mice (C57BL/6 background) were generated by Cyagen Biosciences (Guangzhou, China) using CRISPR/Cas9-mediated deletion of exon 2 of the Disc1 gene.

Techniques: Infection, Staining, Immunofluorescence, Labeling

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Venn diagram showing the overlap of differentially expressed genes (DEGs) identified through transcriptomic and proteomic analyses of ZIKV-infected HTR8 cells compared to non-infected controls. The overlapping area indicates shared DEGs between the two datasets. b − d HTR8 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 hours ( h ) and then infected with ZIKV at a multiplicity of infection (MOI) of 1. Viral mRNA levels, protein expression and titers were assessed on day 1 (D1) and day 2 (D2) post-infection using qRT-PCR ( b ), Western blot ( c ), and plaque assay ( d ). e − g U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h and infected with ZIKV at a MOI of 1. Viral mRNA levels, protein expression and titers were measured on D1 and D2 post-infection using qRT-PCR ( e ), Western blot ( f ), and plaque assay ( g ). h , i HTR8 cells ( h ) or U251 cells ( i ) were transfected with siRNA targeting DISC1 for 24 h prior to ZIKV infection at a MOI of 1. Viral protein was analyzed on D1 and D2 post-infection using Western blot. j − m HTR8 cells ( j , k ) or U251 cells ( l , m ) were transfected with siRNA targeting DISC1 for 24 h prior to ZIKV infection at a MOI of 1. Viral mRNA levels and viral loads were analyzed on D1 and D2 post-infection using qRT-PCR ( j , l ) and plaque assay ( k , m ). Data shown in ( b , d – e , g , j – m ) are from one representative experiment out of three independent replicates with similar results, each including 3 biological replicates per group ( n = 3). Images in ( c − i ) are from one representative experiment out of three independent replicates with similar results ( n = 3). All the data were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using Student’s unpaired two-tailed t test. Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The HA-tagged DISC1 expression plasmid was constructed by inserting the HA-tagged DISC1 coding sequence (NCBI accession number: NM_001012957.2 ) into the pcDNA vector in Sangon Biotechnology (Shanghai, China).

Techniques: Infection, Transfection, Expressing, Quantitative RT-PCR, Western Blot, Plaque Assay, Two Tailed Test

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Schematic representation of the experiment set up. 6-8-week-old WT or Disc1 KD mice were treated with 2 mg MAR1-5A3 on the day prior to infection and then intraperitoneally (i.p.) inoculated with PBS or 1 × 10 6 PFU of ZIKV. Mouse tissues were collected on D2 and D6 post-infection. b − h Viral titers in plasma ( b ) and uterus ( h ) were determined by plaque assay, and mRNA levels in blood cell ( c ), spleen ( d ), brain ( e ), testis ( f ) and uterus ( g ) were measured by qRT-PCR on D2 and D6 post-infection. i , j Primary cells isolated from WT or Disc1 KD mice were induced to differentiate into macrophages using mouse macrophage colony-stimulating factor. Cells were pre-incubated with 20 μg/mL anti-IFNAR1 antibody MAR1-5A3 for 6 h, followed by infection with ZIKV at a MOI of 1. Viral mRNA levels and titers were measured on D1 and D3 post-infection using qRT-PCR ( i ) and plaque assay ( j ). Data for plasma ( b ), blood cells ( c ), spleen ( d ), and brain ( e ) are one representative experiment out of two independent replicates with similar results, each including 6 mice per group ( n = 6), consisting of 3 males and 3 females. Data for the testis ( f ) are one representative experiment out of two independent replicates with similar results, each including 6 male mice per group ( n = 6). Data for the uterus ( g , h ) are one representative experiment out of two independent replicates with similar results, each including 6 female mice per group ( n = 6). Data shown in ( i , j ) are one representative experiment out of two independent replicates with similar results, each including 4 biological replicates per group ( n = 4). Data shown in ( b , e , j ) did not follow a normal distribution according to the Shapiro-Wilk test, and statistical analysis was performed using two-tailed Mann-Whitney test. Data shown in ( c – d , f – i ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using Student’s unpaired two-tailed t test. Data are presented as means ± SD. Source data are provided as a Source Data file. Figure 2a was created in BioRender. Hl, Z. (2025) https://BioRender.com/bh2c5ad .

Article Snippet: The HA-tagged DISC1 expression plasmid was constructed by inserting the HA-tagged DISC1 coding sequence (NCBI accession number: NM_001012957.2 ) into the pcDNA vector in Sangon Biotechnology (Shanghai, China).

Techniques: Infection, Clinical Proteomics, Plaque Assay, Quantitative RT-PCR, Isolation, Incubation, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Schematic representation of the experimental setup. One day prior to infection, 2 mg of the anti-IFNAR1 antibody MAR1-5A3 was administered to 8-10-week-old WT dams and Disc1 KD dams on E5.5. On E6.5, dams were i.p. inoculated with either PBS or 1 × 10 7 PFU of ZIKV. Placenta and fetal head were harvested on E13.5. b Representative images of E13.5 uteri (upper panel) and fetuses (lower panel). Partial demise and growth restriction was shown in ZIKV-infected WT and Disc1 KD pregnant dams. Red arrows indicate the placental residues, and green arrows show the growth restriction of fetuses. c Resorption rates were analyzed on E13.5. d− g The weight ( d ), CRL ( e ), OFD ( f ) and size ( g , CRL × OFD) of fetuses were measured on E13.5. h , i ZIKV mRNA levels in placentas ( h ) and fetal heads ( i ) of WT and Disc1 KD mice was measured by qRT-PCR. j ZIKV mRNA levels in the spleen of in WT or Disc1 KD dams were measured by qRT-PCR. Data shown in ( d − i ) are one representative experiment out of two independent replicates with similar results, each including 4 pregnant dams per group ( n = 4). The offspring analyzed were NF-WT ( n = 36), NF-KD ( n = 37), ZIKV-WT ( n = 28), and ZIKV-KD ( n = 20). Data shown in j are one representative experiment out of two independent replicates with similar results, each including 4 pregnant dams per group ( n = 4). Data shown in ( d − i ) did not follow a normal distribution according to the Shapiro-Wilk test, and statistical analysis was performed using two-tailed Kruskal-Wallis test followed by Dunn’s multiple comparison test ( d − g ) or two-tailed Mann-Whitney test ( h , i ). Data shown in e and j were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test ( e ) or Student’s unpaired two-tailed t test ( j ). Data are presented as means ± SD. Source data are provided as a Source Data file. Figure 3a was created in BioRender. Hl, Z. (2025) https://BioRender.com/dvzm7vs .

Article Snippet: The HA-tagged DISC1 expression plasmid was constructed by inserting the HA-tagged DISC1 coding sequence (NCBI accession number: NM_001012957.2 ) into the pcDNA vector in Sangon Biotechnology (Shanghai, China).

Techniques: Infection, Quantitative RT-PCR, Two Tailed Test, Comparison, MANN-WHITNEY

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Schematic representation of the experiment set up. On the second day after birth, WT and Disc1 KD neonatal mice were intracranially injected with 200 PFU of ZIKV or an equal volume of PBS. Brains were collected on D3, D6 and D9 post-infection, as well as at 6 weeks of age. b ZIKV mRNA levels in WT and Disc1 KD brains were assessed by qRT-PCR on D3, D6 and D9 post-infection, as well as at 6 weeks of age. c , d Body weight ( c ) and brain weight ( d ) of WT and Disc1 KD mice were measured on D3, D6, D9 post-infection, as well as at 6 weeks of age. e , f Representative images of WT and Disc1 KD mice brains on D3, D6, D9 post-infection ( e ), as well as at 6 weeks of age ( f ). The yellow scale bar indicates consistent size within the same age group. g The cerebral size of WT and Disc1 KD mice was measured on D3, D6, D9 post-infection, as well as at 6 weeks of age. Data shown in ( b − d , g ) are collected from two independent replicates, including 8 mice per group ( n = 8), consisting of 4 males and 4 females. Images in ( e , f ) are one representative experiment out of two independent replicates with similar results. Data shown in ( b ) did not follow a normal distribution according to the Shapiro-Wilk test, and statistical analysis was performed using two-tailed Mann-Whitney test. Data shown in ( c – d , g ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test. Data are presented as means ± SD. Source data are provided as a Source Data file. Figure 4a was created in BioRender. Hl, Z. (2025) https://BioRender.com/s3be69y .

Article Snippet: The HA-tagged DISC1 expression plasmid was constructed by inserting the HA-tagged DISC1 coding sequence (NCBI accession number: NM_001012957.2 ) into the pcDNA vector in Sangon Biotechnology (Shanghai, China).

Techniques: Injection, Infection, Quantitative RT-PCR, Two Tailed Test, MANN-WHITNEY, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a On the second day after birth, WT and Disc1 KD neonatal mice were intracranially injected with 200 PFU of ZIKV or an equal volume of PBS, and behavioral tests were conducted when the mice reached 6 weeks of age. The spontaneous alternation rate of WT and Disc1 KD mice in the Y-maze test. b The passing time of WT and Disc1 KD mice in the balance beam test. c , d Proportion of entries into the open arms ( c ) and the percentage of time spent in the open arms ( d ) by WT and Disc1 KD mice in the elevated plus maze test. e , f The time of WT and Disc1 KD mice spent on socializing with Stranger 1 ( e ) and Stranger 2 ( f ) during the 3-chamber test. Data shown in ( a − d ) are collected from one independent experiment, each including 8 mice per group ( n = 8), consisting of 4 males and 4 females. Data shown in ( e , f ) are collected from one independent experiment, each including 10 mice per group ( n = 10), consisting of 5 males and 5 females. Data shown in ( a - b , d – f ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test. Data shown in c did not follow a normal distribution according to the Shapiro-Wilk test, statistical analysis was performed using two-tailed Kruskal-Wallis test followed by Dunn’s multiple comparison test. Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The HA-tagged DISC1 expression plasmid was constructed by inserting the HA-tagged DISC1 coding sequence (NCBI accession number: NM_001012957.2 ) into the pcDNA vector in Sangon Biotechnology (Shanghai, China).

Techniques: Injection, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a KEGG pathway enrichment of DEGs in ZIKV-infected U251 cells transfected with HA-DISC1 (1 μg) plasmid compared with the control group transfected with pcDNA. b KEGG pathway enrichment of differentially expressed proteins identified by IP/MS analysis in U251 cells overexpressing HA-DISC1 (5 μg) plasmid compared with the control group transfected with pcDNA. c Chord diagram illustrates the proteins identified in the IP/MS analysis. d U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h followed by ZIKV infection. The protein levels of AMPKα, pAMPKα, mTOR, pmTOR, P62 and LC3A/B were assessed by Western blot at 0, 12, 24, and 36 h post-infection. e U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h, followed by ZIKV infection for 6 h and treated with DMSO, rapamycin (25 μM), and MHY1485 (10 μM) for 24 h. ZIKV E expression was assessed by Western blot. f U251 cells were transfected with pcDNA or HA-DISC1 (1 μg) plasmids for 24 h, followed by ZIKV infection for 6 h and treated with DMSO, autophagosome inhibitor 3-MA (1 mg/ml), and lysosome inhibitor CQ (100 μM) for 24 h. ZIKV E expression was assessed by Western blot. KEGG pathway enrichment analyses in ( a , b ) were performed using a one-tailed Fisher’s exact test. Images in ( d − f ) are one representative experiment out of three independent replicates with similar results ( n = 3). Source data are provided as a Source Data file.

Article Snippet: The HA-tagged DISC1 expression plasmid was constructed by inserting the HA-tagged DISC1 coding sequence (NCBI accession number: NM_001012957.2 ) into the pcDNA vector in Sangon Biotechnology (Shanghai, China).

Techniques: Infection, Transfection, Plasmid Preparation, Control, Protein-Protein interactions, Western Blot, Expressing, One-tailed Test

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Conserved LIR motifs (amino acids 210 FSFI 213) in DISC1 were highlighted in yellow, which were mutated into AAAA in DISC1 mutant. b 293T cells were co-transfected with HA-DISC1 (5 μg) or HA-DISC1 mutant (5 μg) and GFP-LC3 (5 μg) plasmids for 48 h. Cellular lysates were subjected to immunoprecipitation with anti-Flag or anti-HA magnetic beads and Western blot assays using the indicated antibodies. c − e U251 cells were transfected with HA-DISC1 (1 μg) or HA-DISC1 mutant (1 μg) plasmids for 24 h, followed by ZIKV infection for 24 and 48 h. Viral mRNA levels, protein expression and titers were measured on D1 and D2 post-infection by qRT-PCR ( c ), Western blot ( d ), and plaque assays ( e ). Images in ( b , d ) are from one representative experiment out of three independent replicates with similar results ( n = 3). Data shown in ( c , e ) are from one representative experiment out of three independent replicates with similar results, each including 3 biological replicates per group ( n = 3). Data shown in ( c , e ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using two-tailed one-way ANOVA followed by Tukey’s multiple comparison test. Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The HA-tagged DISC1 expression plasmid was constructed by inserting the HA-tagged DISC1 coding sequence (NCBI accession number: NM_001012957.2 ) into the pcDNA vector in Sangon Biotechnology (Shanghai, China).

Techniques: Mutagenesis, Transfection, Immunoprecipitation, Magnetic Beads, Western Blot, Infection, Expressing, Quantitative RT-PCR, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a Primary cortical neurons were isolated from neonatal mice (P0–P1) and induced to differentiate in vitro. On day 7 of differentiation, neurons exhibiting neurite outgrowth were observed under bright-field microscopy. Confocal imaging showed neuronal nuclei stained with DAPI (blue), and endogenous Neun labeled with an anti-Neun antibody (red). Scale bar = 50 μm. b , c WT and Disc1 KD primary cortical neurons were pre-incubated with 20 μg/mL MAR1-5A3 for 6 h, followed by infection with ZIKV at a MOI of 1. Viral mRNA levels and titers were measured on D1 and D2 post-infection using qRT-PCR ( b ) and plaque assay ( c ). d WT and Disc1 KD primary cortical neurons were pre-incubated with 20 μg/mL MAR1-5A3 for 6 h, followed by infection with ZIKV at a MOI of 1. The protein levels of ZIKV E protein, DISC1, AMPKα, pAMPKα, mTOR, pmTOR, P62 and LC3A/B were assessed by Western blot on D1 and D2 post-infection. e, f The ratios of pAMPKα to AMPKα ( e ) and pmTOR to mTOR ( f ) were determined by comparing the corresponding protein band intensities at each time point in Fig. 8d. g − i Quantification of P62 ( g ), LC3A/BⅠ ( h ) and LC3A/BⅡ ( i ) protein levels relative to GAPDH at each time point in Fig. 8d. j , k Placentas ( j ) and fetal heads ( k ) from non-infected and ZIKV-infected WT and Disc1 KD mice were collected on E13.5. The protein levels of DISC1, AMPKα, pAMPKα, mTOR, pmTOR, P62 and LC3A/B were assessed by Western blot. l − o The ratios of pAMPKα to AMPKα ( l, m ) and pmTOR to mTOR ( n , o ) were determined by comparing the corresponding protein band intensities at each time point in Fig. 8j ( l, n ) and Fig. 8k ( m , o ). p − s Quantification of P62 ( p , q ) and LC3A/BⅡ ( r , s ) protein levels relative to GAPDH at each time point in Fig. 8j ( p , r ) and Fig. 8k ( q , s ). Data shown in ( b , c ) are from one representative experiment out of three independent replicates with similar results, each including 3 biological replicates per group ( n = 3). Images in ( a , d , j – k ) are from one representative experiment out of three independent replicates with similar results ( n = 3). Data shown in ( e − i , l − s) are collected from three independent experiments ( n = 3). Data shown in ( b − c , e − i , l − s ) were confirmed to follow a normal distribution using the Shapiro-Wilk test, and statistical analysis was performed using Student’s unpaired two-tailed t test ( b , c ) or two-tailed one-way ANOVA followed by Tukey’s multiple comparison test ( e − i , l − s ). Data are presented as means ± SD. Source data are provided as a Source Data file.

Article Snippet: The HA-tagged DISC1 expression plasmid was constructed by inserting the HA-tagged DISC1 coding sequence (NCBI accession number: NM_001012957.2 ) into the pcDNA vector in Sangon Biotechnology (Shanghai, China).

Techniques: Isolation, In Vitro, Microscopy, Imaging, Staining, Labeling, Incubation, Infection, Quantitative RT-PCR, Plaque Assay, Western Blot, Two Tailed Test, Comparison

Journal: Nature Communications

Article Title: DISC1 Protects Against Zika Virus Infection and Long-Term Neurological Damage Through AMPK-mTOR-Mediated Autophagy

doi: 10.1038/s41467-025-64809-w

Figure Lengend Snippet: a 6-8-weeks-old WT or Disc1 KD mice were treated with 2 mg MAR1-5A3 on the day prior to infection and then intraperitoneally (i.p.) inoculated with PBS or 1 × 10 6 PFU of ZIKV. Mouse brains were collected on D6 post-infection. Representative H&E-stained sections of the hippocampal DG and CA3 regions are shown. Black arrows indicate shrunken, hyperchromatic neurons with indistinct boundaries Scale bar = 100 μm. b Representative immunofluorescence-stained sections of the hippocampal DG region. Cell nuclei were stained using DAPI (blue). Endogenous Neun was labeled with anti-Neun antibody (red). Endogenous LC3A/B was labeled with anti-LC3A/B antibody (yellow). Scale bar = 100 μm. Representative images in ( a , b ) are from one independent experiment, including 6 mice per group ( n = 6), consisting of 3 males and 3 females.

Article Snippet: The HA-tagged DISC1 expression plasmid was constructed by inserting the HA-tagged DISC1 coding sequence (NCBI accession number: NM_001012957.2 ) into the pcDNA vector in Sangon Biotechnology (Shanghai, China).

Techniques: Infection, Staining, Immunofluorescence, Labeling