Journal: bioRxiv

Article Title: Isoform and protein region abnormalities of dysbindin and copper transporter proteins in postmortem schizophrenia substantia nigra

doi: 10.1101/343178

Figure Lengend Snippet: Schematic of copper transport between the blood and brain in controls (A) and schizophrenia (B). Stars show copper. CTR1, white arrows; ATP7A, black arrows; ATP7B, striped arrows; BBB, blood brain barrier; BCB, blood cerebrospinal barrier. Thinner arrows indicate decreased protein levels.

Article Snippet: The following antibodies and concentrations were used: rabbit anti-N-terminal ATP7A, 1:500, Novus Biologicals (NBP1-54906); rabbit anti-C-terminal ATP7A, 1:2000, Aviva Systems Biology (ARP33798_P050); rabbit anti-ATP7B, 1:1000, Abcam (ab135571); rabbit anti-transmembrane CTR1, 1:2,000, Novus Biologicals (NB100-402); rabbit anti-extracellular CTR1, 1:1000, Aviva Systems Biology (ARP43824_P050); rabbit anti-Dysbindin (targeting 1A and 1B/C), 1:2,000, Abcam (ab133652); and mouse anti-actin, 1:40,000, Millipore (MAB1501).

Techniques:

Journal: bioRxiv

Article Title: Isoform and protein region abnormalities of dysbindin and copper transporter proteins in postmortem schizophrenia substantia nigra

doi: 10.1101/343178

Figure Lengend Snippet: Protein levels of ATP7A (A-B), CTR1 (C-D), dysbindin 1A, 1B/C (E-F) and ATP7B (G-H) for analysis of diagnostic group and treatment status. Error bars represent standard deviation. Significant omnibus ANOVA results are: N-terminal ATP7A: p=0.02; C-terminal ATP7A: p=0.01; transmembrane CTR1: p=0.001. Significant ANOVAs were followed by post hoc tests illustrated by the following: *: p<0.05; **: p<0.01; ***: p<0.001.

Article Snippet: The following antibodies and concentrations were used: rabbit anti-N-terminal ATP7A, 1:500, Novus Biologicals (NBP1-54906); rabbit anti-C-terminal ATP7A, 1:2000, Aviva Systems Biology (ARP33798_P050); rabbit anti-ATP7B, 1:1000, Abcam (ab135571); rabbit anti-transmembrane CTR1, 1:2,000, Novus Biologicals (NB100-402); rabbit anti-extracellular CTR1, 1:1000, Aviva Systems Biology (ARP43824_P050); rabbit anti-Dysbindin (targeting 1A and 1B/C), 1:2,000, Abcam (ab133652); and mouse anti-actin, 1:40,000, Millipore (MAB1501).

Techniques: Diagnostic Assay, Standard Deviation

Journal: bioRxiv

Article Title: Isoform and protein region abnormalities of dysbindin and copper transporter proteins in postmortem schizophrenia substantia nigra

doi: 10.1101/343178

Figure Lengend Snippet: Numerical labeling of amino acid ranges are shown for each protein segment. Dotted boxes indicate antibody-specific epitopes. A. ATP7A protein structure. N-terminal antibody specifically binds to 225-273aa; C-terminal antibody binds to 1403-1452aa. B. CTR1 protein structure. Extracellular CTR1 antibody binds to 19-68aa; transmembrane CTR1 antibody specifically binds to 140-190aa. C. Intracellular diagram of copper chaperones and enzymes and schizophrenia-related alterations. Copper enters neurons after transport from astrocytes via CTR1 and is immediately bound by MT/ GSH. MT/GSH then delivers copper to chaperone ATOX1 or the TGN where it is delivered to other metalloenzymes (e.g., SCO1 → COX) via ATP7A. The SN exhibits more ATOX1 than other brain regions. SCO1 is a copper-requiring enzyme involved in the last step of the electron transport chain of ATP synthesis. SCO1, MT/GSH, COX, ATP, ATP7A and CTR1 are all downregulated or altered in schizophrenia. Abbreviations: MT, metallothionein; GSH, glutathione; APD, antipsychotic drug; SN, substantia nigra; COX, cytochrome c oxidase. References: 1) The present paper; 2) ; 3) ; 4) ; 5) ; 6) ; ; 7) ; 8) ; 9) ; 10) ; and 11) .

Article Snippet: The following antibodies and concentrations were used: rabbit anti-N-terminal ATP7A, 1:500, Novus Biologicals (NBP1-54906); rabbit anti-C-terminal ATP7A, 1:2000, Aviva Systems Biology (ARP33798_P050); rabbit anti-ATP7B, 1:1000, Abcam (ab135571); rabbit anti-transmembrane CTR1, 1:2,000, Novus Biologicals (NB100-402); rabbit anti-extracellular CTR1, 1:1000, Aviva Systems Biology (ARP43824_P050); rabbit anti-Dysbindin (targeting 1A and 1B/C), 1:2,000, Abcam (ab133652); and mouse anti-actin, 1:40,000, Millipore (MAB1501).

Techniques: Labeling

Journal: Parasites & Vectors

Article Title: Cuproptosis-driven astrocyte reactivity exacerbates experimental cerebral malaria pathogenesis

doi: 10.1186/s13071-025-07107-0

Figure Lengend Snippet: Effects of DSF or TTM treatment on astrocytic cuproptosis markers in ECM mice. A - E Representative immunohistochemical staining images of SLC31A1 (copper influx transporter), ATP7A (copper efflux transporter), FDX1 (key cuproptosis regulator), DLAT, and DLST (lipoylated precursor proteins) in the cerebral cortex captured under light microscopy (400 × magnification). Experimental groups: a naive mice; b DSF-treated uninfected controls; c TTM-treated uninfected controls; d untreated Pb A-infected mice; e DSF-treated infected mice; f TTM-treated infected mice. F Quantification of SLC31A1 + , ATP7A + , FDX1 + , DLAT + , and DLST + cells expressed as number per field. Data were obtained from > 20 fields per tissue slice, with six mice per group and three independent experimental replicates. Values represent mean ± standard deviation. Statistical significance: ** P < 0.01 and * P < 0.05 versus infected control (group d); && P < 0.01 versus naive mice (group a); NS (not significant, P > 0.05) indicates no difference compared with naive mice

Article Snippet: Sections were incubated with primary antibodies at 4 °C for overnight: GFAP (1:3,000; GB11096; Servicebio), Serping1 (1:1,000; GB112165 ; Servicebio), SLC31A1 (1:250; NB100-402; Novus), ATP7A (1:100; MA5-27720; Thermo Fisher), FDX1 (1:200; 12592–1-AP; Proteintech), DLAT (1:1,000; GB113649 ; Servicebio), DLST (1:1,500; GB114020 ; Servicebio), and S100B (1:500; GB15359; Servicebio).

Techniques: Immunohistochemical staining, Staining, Light Microscopy, Infection, Standard Deviation, Control

Journal: Parasites & Vectors

Article Title: Cuproptosis-driven astrocyte reactivity exacerbates experimental cerebral malaria pathogenesis

doi: 10.1186/s13071-025-07107-0

Figure Lengend Snippet: Effects of DSF or TTM treatment on GFAP + -SLC31A1 + and GFAP + -FDX1 + astrocytes in the cerebral cortex of ECM mice. A , B Representative double immunofluorescence staining images of GFAP + -SLC31A1 + and GFAP + -FDX1 + astrocytes in the cerebral cortex captured under fluorescence microscopy (400 × magnification). Experimental groups: a naive mice; b DSF-treated uninfected controls; c TTM-treated uninfected controls; d untreated Pb A-infected mice; e DSF-treated infected mice; f TTM-treated infected mice. GFAP + astrocytes (green fluorescence), SLC31A1 + or FDX1 + cells (red fluorescence), and co-localized GFAP + -SLC31A1 + or GFAP + -FDX1 + astrocytes (merged signal, yellow) are shown. C Quantification of GFAP + -SLC31A1 + and GFAP + -FDX1 + astrocytes expressed as number per field. Data were derived from > 20 fields per tissue slice, with six mice per group and three independent experimental replicates. Values represent mean ± standard deviation. Statistical significance: ** P < 0.01 and * P < 0.05 versus Pb A-infected control mice (group d); && P < 0.01 versus naive mice (group a); NS (not significant, P > 0.05) indicates no difference compared with naive mice

Article Snippet: Sections were incubated with primary antibodies at 4 °C for overnight: GFAP (1:3,000; GB11096; Servicebio), Serping1 (1:1,000; GB112165 ; Servicebio), SLC31A1 (1:250; NB100-402; Novus), ATP7A (1:100; MA5-27720; Thermo Fisher), FDX1 (1:200; 12592–1-AP; Proteintech), DLAT (1:1,000; GB113649 ; Servicebio), DLST (1:1,500; GB114020 ; Servicebio), and S100B (1:500; GB15359; Servicebio).

Techniques: Double Immunofluorescence Staining, Fluorescence, Microscopy, Infection, Derivative Assay, Standard Deviation, Control

Journal: Parasites & Vectors

Article Title: Cuproptosis-driven astrocyte reactivity exacerbates experimental cerebral malaria pathogenesis

doi: 10.1186/s13071-025-07107-0

Figure Lengend Snippet: Effects of DSF-CuCl 2 or TTM-CuCl 2 coculture on mRNA expression in iRBCs-stimulated astrocytes in vitro. Total RNA was isolated from astrocytes after coculture under the following conditions: a PBS-treated control (24 h or 48 h); b nonparasitized red blood cells (RBCs)-treated astrocytes (24 h or 48 h); c iRBCs-stimulated astrocytes (24 h or 48 h); d iRBCs-stimulated astrocytes cocultured with DSF-CuCl 2 (24 h or 48 h); e iRBCs-stimulated astrocytes cocultured with TTM-CuCl 2 (24 h or 48 h). mRNA levels of GFAP, Serping1, CXCL10, TNF-α, IL-1β, IL-6, SLC31A1, ATP7A, FDX1, DLAT, and DLST were quantified by qPCR using the 2 −ΔΔCT method. Data were derived from three independent experiments and expressed as mean ± standard deviation. Statistical significance: & P < 0.05 and && P < 0.01, or § P < 0.05 and §§ P < 0.01 versus PBS-treated astrocytes at 24 or 48 h, respectively; * P < 0.05 and ** P < 0.01, or # P < 0.05 and ## P < 0.01 versus iRBCs-stimulated astrocytes at 24 or 48 h, respectively; NS (not significant, P > 0.05) or ns (not significant, P > 0.05) indicates no difference compared with PBS-stimulated astrocytes at 24 or 48 h, respectively

Article Snippet: Sections were incubated with primary antibodies at 4 °C for overnight: GFAP (1:3,000; GB11096; Servicebio), Serping1 (1:1,000; GB112165 ; Servicebio), SLC31A1 (1:250; NB100-402; Novus), ATP7A (1:100; MA5-27720; Thermo Fisher), FDX1 (1:200; 12592–1-AP; Proteintech), DLAT (1:1,000; GB113649 ; Servicebio), DLST (1:1,500; GB114020 ; Servicebio), and S100B (1:500; GB15359; Servicebio).

Techniques: Expressing, In Vitro, Isolation, Control, Derivative Assay, Standard Deviation

Journal: Nature Communications

Article Title: Inflammation mobilizes copper metabolism to promote colon tumorigenesis via an IL-17-STEAP4-XIAP axis

doi: 10.1038/s41467-020-14698-y

Figure Lengend Snippet: a Western blots analysis for copper trafficking-associated proteins from lysates of mouse colon epithelial organoids treated with different inflammatory cytokines for 8 h. Three independent experiments were done and representative blots were shown. Untr untreated. b IL-17 primed or unprimed mouse colon organoids were stained for STEAP4 (St4), E-cadherin (E-cad) and Ctr1 and integrin α. Scale bar, 100 µm. c SOD enzyme activity recovery from TTM inhibition in wild-type (WT) mouse organoids with or without 12 h of IL-17 priming. d SOD enzyme activity recovery in Steap4 wild-type (WT) and knockout (KO) organoids. e , f Fluorometric detection of intracellular copper in primary mouse colon organoids ( e ) or Ls174t cells ( f ). CF4 copper Fluor-4 probe, Ctrl CF4 control probe. Scale bar,100 µm. n = 5 biologically independent cell cultures. g Fluorometric detection of intracellular copper in Steap4 WT and Ctr1 knockdown cells. Cells were treated with or without IL-17 for 12 h and copper were supplemented into the medium for 6 h in all the groups. Scale bar,100 µm. n = 5 biologically independent cell cultures. h Atomic absorption spectroscopy measurement of copper content in Steap4 WT and KO Ls174t cells after copper addition. Cells were primed with or without IL-17 for 12 h and then 10 µM Cu(II) were supplemented into the medium of the indicated group for 6 h. n = 5 biologically independent cell cultures. i Copper content in cytosolic and mitochondrial fraction in Steap4 WT and KO cells, respectively. Cells were treated the same as described in panel ( h ). n = 5 biologically independent cell cultures. Results were normalized to cell numbers. All data show mean ± SEM. Two-tailed Student’s t test was performed for panel e (** P = 0.0057; *** P = 0.0004; **** P < 0.0001), f (*** P = 0.0004; **** P < 0.0001), g (* P = 0.015; ** P = 0.0027; *** P = 0.0004), h (* P = 0.0383; ** P = 0.0022; *** P = 0.0009), i (*** P = 0.0003; **** P < 0.0001; n.s., not significant). Data represent three independent experiments with similar results.

Article Snippet: The following primary antibodies were used for staining presented in the study: Ki67 (Cell Signaling Technology 12202, 1:200), Cleaved Caspase 3 (Cell Signaling Technology 9661, 1:300), STEAP4 (Proteintech, 11944-1-AP,1:100), IL-17A (R&D Systems, MAB317, 1:300), DYKDDDDK (Cell Signaling, 14793S, 1:300), Ctr1 (Novus Biologicals NBP2-36573, 1:100), TUNEL assay (Sigma, 11684795910).

Techniques: Western Blot, Staining, Activity Assay, Inhibition, Knock-Out, Control, Knockdown, Atomic Absorption Spectroscopy, Two Tailed Test