Review



tusc3  (Proteintech)


Bioz Verified Symbol Proteintech is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 93
      Buy from Supplier

    Structured Review

    Proteintech tusc3
    a Schematic of the ER-targeted KDEL-Mag-FRET ER system to assess ER Mg²⁺ dynamics. Created with BioRender.com. b–d SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Circle 1 indicates ER Mg²⁺ release, and Circle 2 represents ER Mg²⁺ refilling (b, left). Representative confocal images at 21 s (b, right). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 6, 7, 7 cells per group) ( d ). e Representative confocal images of SH-SY5Y shTUSC3#3-1 cells co-expressing Mag-FRET ER and <t>TUSC3-mRFP.</t> Scale bar: 10 μm. f – i Non-ID and ID fibroblasts were transfected with Mag-FRET ER alone ( f ) or co-transfected with <t>TUSC3-mRFP</t> ( g ) for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Quantification of normalized ER Mg²⁺ release (Circle 1) ( h ), ER Mg²⁺ uptake ( i , left), and uptake rate ( i , right) (Circle 2) ( n = 7 cells per group). Scale bar: 10 μm. j , k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h, and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( j ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 12, 14 cells per group) ( k ). l , m WT and TUSC3 KO primary cortical neurons were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following acute MgCl₂ treatment (10 mM, 200 s) ( l ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 9 cells per group) ( m ). n , o Non-ID and ID fibroblasts were transfected with Mag-FRET ER for 48 h and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( n ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 7, 10 cells per group) ( o ). One-way ANOVA followed by Tukey’s post hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.
    Tusc3, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tusc3/product/Proteintech
    Average 93 stars, based on 10 article reviews
    tusc3 - by Bioz Stars, 2026-03
    93/100 stars

    Images

    1) Product Images from "TUSC3 regulates ERMA-mediated Mg 2+ uptake for synaptic function and neurodevelopment"

    Article Title: TUSC3 regulates ERMA-mediated Mg 2+ uptake for synaptic function and neurodevelopment

    Journal: Nature Communications

    doi: 10.1038/s41467-025-65668-1

    a Schematic of the ER-targeted KDEL-Mag-FRET ER system to assess ER Mg²⁺ dynamics. Created with BioRender.com. b–d SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Circle 1 indicates ER Mg²⁺ release, and Circle 2 represents ER Mg²⁺ refilling (b, left). Representative confocal images at 21 s (b, right). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 6, 7, 7 cells per group) ( d ). e Representative confocal images of SH-SY5Y shTUSC3#3-1 cells co-expressing Mag-FRET ER and TUSC3-mRFP. Scale bar: 10 μm. f – i Non-ID and ID fibroblasts were transfected with Mag-FRET ER alone ( f ) or co-transfected with TUSC3-mRFP ( g ) for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Quantification of normalized ER Mg²⁺ release (Circle 1) ( h ), ER Mg²⁺ uptake ( i , left), and uptake rate ( i , right) (Circle 2) ( n = 7 cells per group). Scale bar: 10 μm. j , k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h, and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( j ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 12, 14 cells per group) ( k ). l , m WT and TUSC3 KO primary cortical neurons were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following acute MgCl₂ treatment (10 mM, 200 s) ( l ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 9 cells per group) ( m ). n , o Non-ID and ID fibroblasts were transfected with Mag-FRET ER for 48 h and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( n ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 7, 10 cells per group) ( o ). One-way ANOVA followed by Tukey’s post hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.
    Figure Legend Snippet: a Schematic of the ER-targeted KDEL-Mag-FRET ER system to assess ER Mg²⁺ dynamics. Created with BioRender.com. b–d SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Circle 1 indicates ER Mg²⁺ release, and Circle 2 represents ER Mg²⁺ refilling (b, left). Representative confocal images at 21 s (b, right). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 6, 7, 7 cells per group) ( d ). e Representative confocal images of SH-SY5Y shTUSC3#3-1 cells co-expressing Mag-FRET ER and TUSC3-mRFP. Scale bar: 10 μm. f – i Non-ID and ID fibroblasts were transfected with Mag-FRET ER alone ( f ) or co-transfected with TUSC3-mRFP ( g ) for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Quantification of normalized ER Mg²⁺ release (Circle 1) ( h ), ER Mg²⁺ uptake ( i , left), and uptake rate ( i , right) (Circle 2) ( n = 7 cells per group). Scale bar: 10 μm. j , k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h, and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( j ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 12, 14 cells per group) ( k ). l , m WT and TUSC3 KO primary cortical neurons were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following acute MgCl₂ treatment (10 mM, 200 s) ( l ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 9 cells per group) ( m ). n , o Non-ID and ID fibroblasts were transfected with Mag-FRET ER for 48 h and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( n ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 7, 10 cells per group) ( o ). One-way ANOVA followed by Tukey’s post hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.

    Techniques Used: Transfection, Expressing, Comparison

    a–d SH-SY5Y shControl and shTUSC3#3-1 cells were co-transfected with Mag-FRET ER and ERMA-DsRed for 48 h and analyzed using confocal microscopy ( a ). Scale bar: 10 μm. Representative traces showing ER Mg²⁺ uptake following L-lactate (10 mM) treatment ( b ). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 9 cells per group) ( d ). e , f Co-immunoprecipitation from whole brain lysates of WT mice showing endogenous interaction between TUSC3 and ERMA. Immunoprecipitation with anti-ERMA antibody pulled down TUSC3 ( e ), and reciprocal IP with anti-TUSC3 antibody confirmed the presence of ERMA ( f ). L.C.: light chain of immunoglobulin. g Schematic representation of TUSC3-mRFP deletion mutants. h Immunoprecipitation assays between ERMA-FLAG and various TUSC3-mRFP deletion mutants. i – k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER and indicated TUSC3-mRFP deletion mutants for 48 h and analyzed for ER Mg²⁺ release and uptake following L-lactate (10 mM) treatment ( i ). Quantification of ER Mg²⁺ release (Circle 1) ( j ) and ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( k ) ( n = 10, 9, 9, 6, 9, 4, 5 cells per group). One-way ANOVA followed by Tukey’s or Dunnett’s ( k ) post-hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.
    Figure Legend Snippet: a–d SH-SY5Y shControl and shTUSC3#3-1 cells were co-transfected with Mag-FRET ER and ERMA-DsRed for 48 h and analyzed using confocal microscopy ( a ). Scale bar: 10 μm. Representative traces showing ER Mg²⁺ uptake following L-lactate (10 mM) treatment ( b ). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 9 cells per group) ( d ). e , f Co-immunoprecipitation from whole brain lysates of WT mice showing endogenous interaction between TUSC3 and ERMA. Immunoprecipitation with anti-ERMA antibody pulled down TUSC3 ( e ), and reciprocal IP with anti-TUSC3 antibody confirmed the presence of ERMA ( f ). L.C.: light chain of immunoglobulin. g Schematic representation of TUSC3-mRFP deletion mutants. h Immunoprecipitation assays between ERMA-FLAG and various TUSC3-mRFP deletion mutants. i – k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER and indicated TUSC3-mRFP deletion mutants for 48 h and analyzed for ER Mg²⁺ release and uptake following L-lactate (10 mM) treatment ( i ). Quantification of ER Mg²⁺ release (Circle 1) ( j ) and ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( k ) ( n = 10, 9, 9, 6, 9, 4, 5 cells per group). One-way ANOVA followed by Tukey’s or Dunnett’s ( k ) post-hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.

    Techniques Used: Transfection, Confocal Microscopy, Immunoprecipitation, Comparison

    a Representative western blot of ER stress proteins in the hippocampus of 4-month-old female WT and TUSC3 KO mice. b – i Quantification of GRP78 ( b ), p-PERK (T982) ( c ), p-eIF2α (S51) ( d ), CHOP ( e ), XBP1s ( f ), p-AKT (S473) ( g ), p-CREB (S133) ( h ), and ATF6 ( i ) normalized to β-actin ( n = 3 mice per group). j Representative western blot analysis of ER stress proteins in the striatum of 4-month-old female WT and TUSC3 KO mice ( n = 3 per group). k – r Quantification of GRP78 ( k ), p-PERK (T982) ( l ), p-eIF2α (S51) ( m ), CHOP ( n ), XBP1s ( o ), p-AKT (S473) ( p ), p-CREB (S133) ( q ), and ATF6 ( r ) normalized to β-actin ( n = 3 mice per group). s Non-ID and ID1/ID2 fibroblasts were chronically treated with MgT (400 μM) for 48 h and analyzed by western blotting (left). Quantification of p-eIF2α (S51) levels normalized to eIF2α (right) ( n = 3 independent biological replicates). t Non-ID and ID-derived fibroblasts were pretreated with or without MgT (400 μM) for 24 h, followed by puromycin (5 μg/ml) treatment for 2 h. Representative western blot using an anti-puromycin antibody (left), and puromycin incorporation was quantified and normalized to β-actin (right) ( n = 3 independent biological replicates). u SH-SY5Y shControl and shTUSC3#3-1 cells were pretreated with MgT (400 μM) for 12 h, then treated with either DMSO, thapsigargin (2 μM), tunicamycin (2 μg/ml), A23187 (2 μM), or etoposide (25 μM) for 24 h. Cells were stained with propidium iodide and Calcein-AM, and double-positive cells were quantified and normalized to Calcein-AM-positive cells. Data represent three independent biological experiments ( n = 3). The exact number of cells analyzed per condition is provided in the Source Data file. v Representative western blot of primary cortical neurons (DIV7) treated with vehicle or 4-PBA (10 mM) for 24 h (left). Quantification of indicated protein levels normalized to TUBA (right) ( n = 3 independent biological replicates). Two-tailed unpaired t -test ( b – i , and k – r ); one-way ANOVA followed by Tukey’s post hoc multiple comparison test ( s , t , v ); two-way ANOVA followed by Tukey’s multiple comparison test ( u ). Data are presented as mean ± S.E.M. or S.D. ( t , u ). Source data are provided as a file.
    Figure Legend Snippet: a Representative western blot of ER stress proteins in the hippocampus of 4-month-old female WT and TUSC3 KO mice. b – i Quantification of GRP78 ( b ), p-PERK (T982) ( c ), p-eIF2α (S51) ( d ), CHOP ( e ), XBP1s ( f ), p-AKT (S473) ( g ), p-CREB (S133) ( h ), and ATF6 ( i ) normalized to β-actin ( n = 3 mice per group). j Representative western blot analysis of ER stress proteins in the striatum of 4-month-old female WT and TUSC3 KO mice ( n = 3 per group). k – r Quantification of GRP78 ( k ), p-PERK (T982) ( l ), p-eIF2α (S51) ( m ), CHOP ( n ), XBP1s ( o ), p-AKT (S473) ( p ), p-CREB (S133) ( q ), and ATF6 ( r ) normalized to β-actin ( n = 3 mice per group). s Non-ID and ID1/ID2 fibroblasts were chronically treated with MgT (400 μM) for 48 h and analyzed by western blotting (left). Quantification of p-eIF2α (S51) levels normalized to eIF2α (right) ( n = 3 independent biological replicates). t Non-ID and ID-derived fibroblasts were pretreated with or without MgT (400 μM) for 24 h, followed by puromycin (5 μg/ml) treatment for 2 h. Representative western blot using an anti-puromycin antibody (left), and puromycin incorporation was quantified and normalized to β-actin (right) ( n = 3 independent biological replicates). u SH-SY5Y shControl and shTUSC3#3-1 cells were pretreated with MgT (400 μM) for 12 h, then treated with either DMSO, thapsigargin (2 μM), tunicamycin (2 μg/ml), A23187 (2 μM), or etoposide (25 μM) for 24 h. Cells were stained with propidium iodide and Calcein-AM, and double-positive cells were quantified and normalized to Calcein-AM-positive cells. Data represent three independent biological experiments ( n = 3). The exact number of cells analyzed per condition is provided in the Source Data file. v Representative western blot of primary cortical neurons (DIV7) treated with vehicle or 4-PBA (10 mM) for 24 h (left). Quantification of indicated protein levels normalized to TUBA (right) ( n = 3 independent biological replicates). Two-tailed unpaired t -test ( b – i , and k – r ); one-way ANOVA followed by Tukey’s post hoc multiple comparison test ( s , t , v ); two-way ANOVA followed by Tukey’s multiple comparison test ( u ). Data are presented as mean ± S.E.M. or S.D. ( t , u ). Source data are provided as a file.

    Techniques Used: Western Blot, Derivative Assay, Staining, Two Tailed Test, Comparison

    a , b Schematic of the Y-maze test ( a ) and spatial memory performance in WT and TUSC3 KO mice ( b ). Created with BioRender.com. Schematic of the novel object recognition (NOR) test ( c ) and representative tracking plots in the open field in the NOR test ( d ). Created with BioRender.com. e The NOR test (discrimination index) for WT and TUSC3 KO mice. f , g Schematic of the forced swim test (FST), consisting of a 120 s adaptation phase followed by 240 s of immobility measurement ( f ) and immobility time in the FST, assessing stress-coping ability in WT and TUSC3 KO mice ( g ). Created with BioRender.com. h , i Schematic of the tail suspension test (TST), with immobility recorded over 360 s ( h ) and immobility time in the TST for WT and TUSC3 KO mice ( i ). Created with BioRender.com. j , k Schematic of three-chambered social interaction test ( j ) and representative tracking plots from the social interaction test, showing times spent in each chamber ( k ). Created with BioRender.com. l , m Social interaction preference in TUSC3 KO mice, assessed by time spent with the novel mouse ( l ) and sniffing time ( m ). n – q Schematic of three-chambered social preference test ( n ) and representative tracking plots of WT and TUSC3 KO mice in social preference test ( o ). Social preference for the novel mouse of WT and TUSC3 KO mice in the social preference test ( p ). Sniffing time data confirms this lack of preference ( q ). Created with BioRender.com. In all behavioral experiments, 4-month-old mice were used: WT: 9 (5 F, 4 M); KO: 10 (6 F, 4 M). Two-tailed unpaired t -test ( b , e , g , i ), Two-way ANOVA followed by Tukey’s post-hoc multiple comparison test ( l , m , p , q ). All data are presented as mean ± S.E.M. Source data are provided as a file.
    Figure Legend Snippet: a , b Schematic of the Y-maze test ( a ) and spatial memory performance in WT and TUSC3 KO mice ( b ). Created with BioRender.com. Schematic of the novel object recognition (NOR) test ( c ) and representative tracking plots in the open field in the NOR test ( d ). Created with BioRender.com. e The NOR test (discrimination index) for WT and TUSC3 KO mice. f , g Schematic of the forced swim test (FST), consisting of a 120 s adaptation phase followed by 240 s of immobility measurement ( f ) and immobility time in the FST, assessing stress-coping ability in WT and TUSC3 KO mice ( g ). Created with BioRender.com. h , i Schematic of the tail suspension test (TST), with immobility recorded over 360 s ( h ) and immobility time in the TST for WT and TUSC3 KO mice ( i ). Created with BioRender.com. j , k Schematic of three-chambered social interaction test ( j ) and representative tracking plots from the social interaction test, showing times spent in each chamber ( k ). Created with BioRender.com. l , m Social interaction preference in TUSC3 KO mice, assessed by time spent with the novel mouse ( l ) and sniffing time ( m ). n – q Schematic of three-chambered social preference test ( n ) and representative tracking plots of WT and TUSC3 KO mice in social preference test ( o ). Social preference for the novel mouse of WT and TUSC3 KO mice in the social preference test ( p ). Sniffing time data confirms this lack of preference ( q ). Created with BioRender.com. In all behavioral experiments, 4-month-old mice were used: WT: 9 (5 F, 4 M); KO: 10 (6 F, 4 M). Two-tailed unpaired t -test ( b , e , g , i ), Two-way ANOVA followed by Tukey’s post-hoc multiple comparison test ( l , m , p , q ). All data are presented as mean ± S.E.M. Source data are provided as a file.

    Techniques Used: Suspension, Two Tailed Test, Comparison

    a Representative western blot of PSD-93, PSD-95, GluA1, and phosphorylated CREB (S133) in whole-brain lysates of 4-month-old WT and TUSC3 KO female mice (left). Quantification of indicated protein levels (right) ( n = 3). b , c Representative confocal images of immunohistochemistry for TUSC3, synaptophysin (SYP), and MAP2 in the CA1 and CA3 hippocampal regions and the striatum of 4-month-old WT and TUSC3 KO female mice ( b ), and quantification of SYP intensity normalized to MAP2 ( c ) ( n = 3). Scale bar: 50 μm. d , e Representative confocal images of immunohistochemistry for TUSC3, GluA1, and MAP2 in the CA1 and CA3 hippocampal regions and the striatum of 4-month-old WT and TUSC3 KO female mice ( d ), and quantification of GluA1 intensity normalized to MAP2 ( e ) ( n = 3). Scale bar: 50 μm. f , g Representative dendritic images of primary hippocampal neurons transfected with EGFP ( f ). Scale bar: 50 μm. Sholl analysis of dendritic branching in WT and TUSC3 KO neurons ( n = 20 cells per group) ( g ). h , i Representative confocal images of dendritic spines of primary hippocampal neurons (DIV 12) transfected with EGFP for 48 h ( h ). Violin plot quantifying dendritic spine density (spines per 10 μm) in WT and KO neurons ( n = 20 cells per group) ( i ). Scale bar: 10 μm. j–l Representative traces of sEPSCs recorded from hippocampal slices of 2-month-old WT and TUSC3 KO mice ( j ), quantification of sEPSC amplitude ( k ), and frequency ( l ) in WT ( n = 12 cells from 5 mice; 4 F, 1 M) and TUSC3 KO ( n = 8 cells from 5 mice; 2 F, 3 M). m Representative traces (left) and quantification of the eEPSC amplitude (right) in WT ( n = 8 cells from 5 mice; 4 F, 1 M), TUSC3 KO ( n = 8 cells from 5 mice; 2 F, 3 M). n Representative traces (left) and EPSC-PPR (right) in WT ( n = 8 cells from 5 mice; 3 F, 2 M), TUSC3 KO ( n = 9 cells from 6 mice; 4 F, 2 M). Two-tailed unpaired t -test. Data are presented as mean ± S.E.M. ( a , c , e , and k – n ) or S.D. ( g , i ). Source data are provided as a file.
    Figure Legend Snippet: a Representative western blot of PSD-93, PSD-95, GluA1, and phosphorylated CREB (S133) in whole-brain lysates of 4-month-old WT and TUSC3 KO female mice (left). Quantification of indicated protein levels (right) ( n = 3). b , c Representative confocal images of immunohistochemistry for TUSC3, synaptophysin (SYP), and MAP2 in the CA1 and CA3 hippocampal regions and the striatum of 4-month-old WT and TUSC3 KO female mice ( b ), and quantification of SYP intensity normalized to MAP2 ( c ) ( n = 3). Scale bar: 50 μm. d , e Representative confocal images of immunohistochemistry for TUSC3, GluA1, and MAP2 in the CA1 and CA3 hippocampal regions and the striatum of 4-month-old WT and TUSC3 KO female mice ( d ), and quantification of GluA1 intensity normalized to MAP2 ( e ) ( n = 3). Scale bar: 50 μm. f , g Representative dendritic images of primary hippocampal neurons transfected with EGFP ( f ). Scale bar: 50 μm. Sholl analysis of dendritic branching in WT and TUSC3 KO neurons ( n = 20 cells per group) ( g ). h , i Representative confocal images of dendritic spines of primary hippocampal neurons (DIV 12) transfected with EGFP for 48 h ( h ). Violin plot quantifying dendritic spine density (spines per 10 μm) in WT and KO neurons ( n = 20 cells per group) ( i ). Scale bar: 10 μm. j–l Representative traces of sEPSCs recorded from hippocampal slices of 2-month-old WT and TUSC3 KO mice ( j ), quantification of sEPSC amplitude ( k ), and frequency ( l ) in WT ( n = 12 cells from 5 mice; 4 F, 1 M) and TUSC3 KO ( n = 8 cells from 5 mice; 2 F, 3 M). m Representative traces (left) and quantification of the eEPSC amplitude (right) in WT ( n = 8 cells from 5 mice; 4 F, 1 M), TUSC3 KO ( n = 8 cells from 5 mice; 2 F, 3 M). n Representative traces (left) and EPSC-PPR (right) in WT ( n = 8 cells from 5 mice; 3 F, 2 M), TUSC3 KO ( n = 9 cells from 6 mice; 4 F, 2 M). Two-tailed unpaired t -test. Data are presented as mean ± S.E.M. ( a , c , e , and k – n ) or S.D. ( g , i ). Source data are provided as a file.

    Techniques Used: Western Blot, Immunohistochemistry, Transfection, Two Tailed Test

    a Experimental timeline. Four-week-old mice received chronic magnesium supplementation via drinking water containing MgT (910 mg/kg/day) for 42 days, followed by behavioral analysis. Created with BioRender.com. b , c Cognitive performance of MgT-treated WT and TUSC3 KO mice assessed in the Y-maze ( b ) and NOR test ( c ). d Stress-coping ability evaluated using the TST. e , f Social behavior assessed in the social interaction ( e ) and social novelty preference ( f ) tests (left). Time spent in each chamber (middle) and sniffing time (right) were measured. g , h Representative western blot of hippocampal tissues from WT and TUSC3 KO mice treated with vehicle or MgT ( g ). Quantification of indicated protein levels ( h ) ( n = 3). i , j Representative confocal images of GluA1 immunohistochemistry in the CA1 ( i ) and CA3 ( j ) hippocampal regions from WT and TUSC3 KO mice treated with vehicle or MgT ( n = 3). Scale bar: 50 μm. In these experiments, 10-week-old mice were used: WT: 8 (4 F, 4 M); WT + MgT: 8 (4 F, 4 M); KO: 6 (3 F, 3 M); KO + MgT: 8 (5 F, 3 M). Two-way ANOVA with Tukey’s post hoc multiple comparison test. All data are represented as mean ± S.E.M. Source data are provided as a file.
    Figure Legend Snippet: a Experimental timeline. Four-week-old mice received chronic magnesium supplementation via drinking water containing MgT (910 mg/kg/day) for 42 days, followed by behavioral analysis. Created with BioRender.com. b , c Cognitive performance of MgT-treated WT and TUSC3 KO mice assessed in the Y-maze ( b ) and NOR test ( c ). d Stress-coping ability evaluated using the TST. e , f Social behavior assessed in the social interaction ( e ) and social novelty preference ( f ) tests (left). Time spent in each chamber (middle) and sniffing time (right) were measured. g , h Representative western blot of hippocampal tissues from WT and TUSC3 KO mice treated with vehicle or MgT ( g ). Quantification of indicated protein levels ( h ) ( n = 3). i , j Representative confocal images of GluA1 immunohistochemistry in the CA1 ( i ) and CA3 ( j ) hippocampal regions from WT and TUSC3 KO mice treated with vehicle or MgT ( n = 3). Scale bar: 50 μm. In these experiments, 10-week-old mice were used: WT: 8 (4 F, 4 M); WT + MgT: 8 (4 F, 4 M); KO: 6 (3 F, 3 M); KO + MgT: 8 (5 F, 3 M). Two-way ANOVA with Tukey’s post hoc multiple comparison test. All data are represented as mean ± S.E.M. Source data are provided as a file.

    Techniques Used: Western Blot, Immunohistochemistry, Comparison

    TUSC3 deficiency impairs ER magnesium uptake by disrupting its positive modulation of ERMA, an ER Mg²⁺ transporter. This leads to reduced ER [Mg²⁺], activation of ER stress via the PERK–eIF2α axis, and suppression of global translation, including synaptic proteins (PSD-93, PSD-95, GluA1). In parallel, CREB activity is reduced through impaired PI3K/AKT signaling. Together, these changes result in synaptic dysfunction, increased neuronal vulnerability (via CHOP), and ultimately cognitive and adaptive behavioral deficits. Notably, magnesium supplementation restores ER [Mg²⁺] levels, alleviates ER stress, and rescues synaptic and cognitive phenotypes in TUSC3-deficient mice, suggesting a potential therapeutic strategy for TUSC3-associated ID.
    Figure Legend Snippet: TUSC3 deficiency impairs ER magnesium uptake by disrupting its positive modulation of ERMA, an ER Mg²⁺ transporter. This leads to reduced ER [Mg²⁺], activation of ER stress via the PERK–eIF2α axis, and suppression of global translation, including synaptic proteins (PSD-93, PSD-95, GluA1). In parallel, CREB activity is reduced through impaired PI3K/AKT signaling. Together, these changes result in synaptic dysfunction, increased neuronal vulnerability (via CHOP), and ultimately cognitive and adaptive behavioral deficits. Notably, magnesium supplementation restores ER [Mg²⁺] levels, alleviates ER stress, and rescues synaptic and cognitive phenotypes in TUSC3-deficient mice, suggesting a potential therapeutic strategy for TUSC3-associated ID.

    Techniques Used: Activation Assay, Activity Assay



    Similar Products

    tusc3  (Proteintech)
    93
    Proteintech tusc3
    a Schematic of the ER-targeted KDEL-Mag-FRET ER system to assess ER Mg²⁺ dynamics. Created with BioRender.com. b–d SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Circle 1 indicates ER Mg²⁺ release, and Circle 2 represents ER Mg²⁺ refilling (b, left). Representative confocal images at 21 s (b, right). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 6, 7, 7 cells per group) ( d ). e Representative confocal images of SH-SY5Y shTUSC3#3-1 cells co-expressing Mag-FRET ER and <t>TUSC3-mRFP.</t> Scale bar: 10 μm. f – i Non-ID and ID fibroblasts were transfected with Mag-FRET ER alone ( f ) or co-transfected with <t>TUSC3-mRFP</t> ( g ) for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Quantification of normalized ER Mg²⁺ release (Circle 1) ( h ), ER Mg²⁺ uptake ( i , left), and uptake rate ( i , right) (Circle 2) ( n = 7 cells per group). Scale bar: 10 μm. j , k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h, and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( j ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 12, 14 cells per group) ( k ). l , m WT and TUSC3 KO primary cortical neurons were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following acute MgCl₂ treatment (10 mM, 200 s) ( l ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 9 cells per group) ( m ). n , o Non-ID and ID fibroblasts were transfected with Mag-FRET ER for 48 h and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( n ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 7, 10 cells per group) ( o ). One-way ANOVA followed by Tukey’s post hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.
    Tusc3, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tusc3/product/Proteintech
    Average 93 stars, based on 1 article reviews
    tusc3 - by Bioz Stars, 2026-03
    93/100 stars
    		  Buy from Supplier
    antibodies against tusc3  (Proteintech)
    93
    Proteintech antibodies against tusc3
    a Schematic of the ER-targeted KDEL-Mag-FRET ER system to assess ER Mg²⁺ dynamics. Created with BioRender.com. b–d SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Circle 1 indicates ER Mg²⁺ release, and Circle 2 represents ER Mg²⁺ refilling (b, left). Representative confocal images at 21 s (b, right). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 6, 7, 7 cells per group) ( d ). e Representative confocal images of SH-SY5Y shTUSC3#3-1 cells co-expressing Mag-FRET ER and <t>TUSC3-mRFP.</t> Scale bar: 10 μm. f – i Non-ID and ID fibroblasts were transfected with Mag-FRET ER alone ( f ) or co-transfected with <t>TUSC3-mRFP</t> ( g ) for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Quantification of normalized ER Mg²⁺ release (Circle 1) ( h ), ER Mg²⁺ uptake ( i , left), and uptake rate ( i , right) (Circle 2) ( n = 7 cells per group). Scale bar: 10 μm. j , k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h, and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( j ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 12, 14 cells per group) ( k ). l , m WT and TUSC3 KO primary cortical neurons were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following acute MgCl₂ treatment (10 mM, 200 s) ( l ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 9 cells per group) ( m ). n , o Non-ID and ID fibroblasts were transfected with Mag-FRET ER for 48 h and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( n ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 7, 10 cells per group) ( o ). One-way ANOVA followed by Tukey’s post hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.
    Antibodies Against Tusc3, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/antibodies against tusc3/product/Proteintech
    Average 93 stars, based on 1 article reviews
    antibodies against tusc3 - by Bioz Stars, 2026-03
    93/100 stars
    		  Buy from Supplier
    tusc3 67382 antibody  (Proteintech)
    90
    Proteintech tusc3 67382 antibody
    a Schematic of the ER-targeted KDEL-Mag-FRET ER system to assess ER Mg²⁺ dynamics. Created with BioRender.com. b–d SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Circle 1 indicates ER Mg²⁺ release, and Circle 2 represents ER Mg²⁺ refilling (b, left). Representative confocal images at 21 s (b, right). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 6, 7, 7 cells per group) ( d ). e Representative confocal images of SH-SY5Y shTUSC3#3-1 cells co-expressing Mag-FRET ER and <t>TUSC3-mRFP.</t> Scale bar: 10 μm. f – i Non-ID and ID fibroblasts were transfected with Mag-FRET ER alone ( f ) or co-transfected with <t>TUSC3-mRFP</t> ( g ) for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Quantification of normalized ER Mg²⁺ release (Circle 1) ( h ), ER Mg²⁺ uptake ( i , left), and uptake rate ( i , right) (Circle 2) ( n = 7 cells per group). Scale bar: 10 μm. j , k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h, and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( j ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 12, 14 cells per group) ( k ). l , m WT and TUSC3 KO primary cortical neurons were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following acute MgCl₂ treatment (10 mM, 200 s) ( l ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 9 cells per group) ( m ). n , o Non-ID and ID fibroblasts were transfected with Mag-FRET ER for 48 h and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( n ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 7, 10 cells per group) ( o ). One-way ANOVA followed by Tukey’s post hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.
    Tusc3 67382 Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/tusc3 67382 antibody/product/Proteintech
    Average 90 stars, based on 1 article reviews
    tusc3 67382 antibody - by Bioz Stars, 2026-03
    90/100 stars
    		  Buy from Supplier

    Image Search Results


    a Schematic of the ER-targeted KDEL-Mag-FRET ER system to assess ER Mg²⁺ dynamics. Created with BioRender.com. b–d SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Circle 1 indicates ER Mg²⁺ release, and Circle 2 represents ER Mg²⁺ refilling (b, left). Representative confocal images at 21 s (b, right). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 6, 7, 7 cells per group) ( d ). e Representative confocal images of SH-SY5Y shTUSC3#3-1 cells co-expressing Mag-FRET ER and TUSC3-mRFP. Scale bar: 10 μm. f – i Non-ID and ID fibroblasts were transfected with Mag-FRET ER alone ( f ) or co-transfected with TUSC3-mRFP ( g ) for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Quantification of normalized ER Mg²⁺ release (Circle 1) ( h ), ER Mg²⁺ uptake ( i , left), and uptake rate ( i , right) (Circle 2) ( n = 7 cells per group). Scale bar: 10 μm. j , k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h, and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( j ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 12, 14 cells per group) ( k ). l , m WT and TUSC3 KO primary cortical neurons were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following acute MgCl₂ treatment (10 mM, 200 s) ( l ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 9 cells per group) ( m ). n , o Non-ID and ID fibroblasts were transfected with Mag-FRET ER for 48 h and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( n ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 7, 10 cells per group) ( o ). One-way ANOVA followed by Tukey’s post hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.

    Journal: Nature Communications

    Article Title: TUSC3 regulates ERMA-mediated Mg 2+ uptake for synaptic function and neurodevelopment

    doi: 10.1038/s41467-025-65668-1

    Figure Lengend Snippet: a Schematic of the ER-targeted KDEL-Mag-FRET ER system to assess ER Mg²⁺ dynamics. Created with BioRender.com. b–d SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Circle 1 indicates ER Mg²⁺ release, and Circle 2 represents ER Mg²⁺ refilling (b, left). Representative confocal images at 21 s (b, right). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 6, 7, 7 cells per group) ( d ). e Representative confocal images of SH-SY5Y shTUSC3#3-1 cells co-expressing Mag-FRET ER and TUSC3-mRFP. Scale bar: 10 μm. f – i Non-ID and ID fibroblasts were transfected with Mag-FRET ER alone ( f ) or co-transfected with TUSC3-mRFP ( g ) for 48 h and analyzed for ER Mg²⁺ uptake following L-lactate treatment (10 mM). Quantification of normalized ER Mg²⁺ release (Circle 1) ( h ), ER Mg²⁺ uptake ( i , left), and uptake rate ( i , right) (Circle 2) ( n = 7 cells per group). Scale bar: 10 μm. j , k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER for 48 h, and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( j ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 12, 14 cells per group) ( k ). l , m WT and TUSC3 KO primary cortical neurons were transfected with Mag-FRET ER for 48 h and analyzed for ER Mg²⁺ uptake following acute MgCl₂ treatment (10 mM, 200 s) ( l ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 9 cells per group) ( m ). n , o Non-ID and ID fibroblasts were transfected with Mag-FRET ER for 48 h and supplemented with acute MgCl₂ treatment (10 mM, 200 s) to assess ER Mg²⁺ uptake ( n ), quantification of ER Mg²⁺ levels at 0 s and 200 s ( n = 10, 7, 10 cells per group) ( o ). One-way ANOVA followed by Tukey’s post hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.

    Article Snippet: The following primary antibodies were used in this study: ACTB (Santa Cruz Biotechnology; sc-47778; lot # J0421; WB 1:5000), TUBA (Santa Cruz Biotechnology; sc-23948; lot # G2921; WB 1:5000), TUSC3 (clone D-9; Santa Cruz Biotechnology; sc-390566; lot # J1218; IF 1:500), GRP78 (Santa Cruz Biotechnology; sc-376768; lot # C0316; WB 1:3000), p-CREB-1-S133 (clone 10E9; Santa Cruz Biotechnology; sc-81486; WB 1:3000), FLAG (Sigma-Aldrich; F1804; lot # 0000375542; WB 1:3000), RFP (MBL; PM005; lot # 048; WB 1:3000), TUSC3 (Proteintech; 16039-1-AP; lot # 00007251; WB 1:500), AKT (Cell Signaling Technology; 9272; lot # 6; WB 1:3000), P-AKT (S473) (Cell Signaling Technology; 4060; lot # 27; WB 1:3000; IF 1:250), CREB (Cell Signaling Technology; 9197; WB 1:1000), CHOP (Cell Signaling Technology; 2895; WB 1:1000), ATF6 (Cell Signaling Technology; 65880; lot # 5; WB 1:3000), MAP2 (Cell Signaling Technology; 4542; IF 1:500), NCAM1 (Cell Signaling Technology; 3576; WB 1:3000), PERK (ABclonal; A18196; lot # 3560572205; WB 1:3000), p-PERK (T982) (ABclonal; AP0886; lot # 5500040024; WB 1:3000), XBP1s (ABclonal; A17007; lot # 1153240401; WB 1:3000), eIF2α (ABclonal; A21221; lot # 360002151; WB 1:3000), p-eIF2α (ABclonal; AP0745; lot # 360003834; WB 1:3000), Puromycin (ABclonal; A23931; lot # 3523033102; WB 1:3000), PSD-93 (ABclonal; A19669; WB 1:3000), PSD-95 (ABclonal; A7889; WB 1:3000), GluA1 (ABclonal; A1826; lot # 3561869003; WB 1:3000; IF 1:250), NMDAR1 (ABclonal; A7677; lot # 5500016744; WB 1:1000), Synaptophysin (ABclonal; A6344; lot # 5500008363; WB 1:1000; IF 1:250), and N-Cadherin (BD Biosciences; 610920; WB 1:3000).

    Techniques: Transfection, Expressing, Comparison

    a–d SH-SY5Y shControl and shTUSC3#3-1 cells were co-transfected with Mag-FRET ER and ERMA-DsRed for 48 h and analyzed using confocal microscopy ( a ). Scale bar: 10 μm. Representative traces showing ER Mg²⁺ uptake following L-lactate (10 mM) treatment ( b ). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 9 cells per group) ( d ). e , f Co-immunoprecipitation from whole brain lysates of WT mice showing endogenous interaction between TUSC3 and ERMA. Immunoprecipitation with anti-ERMA antibody pulled down TUSC3 ( e ), and reciprocal IP with anti-TUSC3 antibody confirmed the presence of ERMA ( f ). L.C.: light chain of immunoglobulin. g Schematic representation of TUSC3-mRFP deletion mutants. h Immunoprecipitation assays between ERMA-FLAG and various TUSC3-mRFP deletion mutants. i – k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER and indicated TUSC3-mRFP deletion mutants for 48 h and analyzed for ER Mg²⁺ release and uptake following L-lactate (10 mM) treatment ( i ). Quantification of ER Mg²⁺ release (Circle 1) ( j ) and ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( k ) ( n = 10, 9, 9, 6, 9, 4, 5 cells per group). One-way ANOVA followed by Tukey’s or Dunnett’s ( k ) post-hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.

    Journal: Nature Communications

    Article Title: TUSC3 regulates ERMA-mediated Mg 2+ uptake for synaptic function and neurodevelopment

    doi: 10.1038/s41467-025-65668-1

    Figure Lengend Snippet: a–d SH-SY5Y shControl and shTUSC3#3-1 cells were co-transfected with Mag-FRET ER and ERMA-DsRed for 48 h and analyzed using confocal microscopy ( a ). Scale bar: 10 μm. Representative traces showing ER Mg²⁺ uptake following L-lactate (10 mM) treatment ( b ). Quantification of normalized ER Mg²⁺ release (Circle 1) ( c ). Quantification of normalized ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( n = 9 cells per group) ( d ). e , f Co-immunoprecipitation from whole brain lysates of WT mice showing endogenous interaction between TUSC3 and ERMA. Immunoprecipitation with anti-ERMA antibody pulled down TUSC3 ( e ), and reciprocal IP with anti-TUSC3 antibody confirmed the presence of ERMA ( f ). L.C.: light chain of immunoglobulin. g Schematic representation of TUSC3-mRFP deletion mutants. h Immunoprecipitation assays between ERMA-FLAG and various TUSC3-mRFP deletion mutants. i – k SH-SY5Y shControl and shTUSC3#3-1 cells were transfected with Mag-FRET ER and indicated TUSC3-mRFP deletion mutants for 48 h and analyzed for ER Mg²⁺ release and uptake following L-lactate (10 mM) treatment ( i ). Quantification of ER Mg²⁺ release (Circle 1) ( j ) and ER Mg²⁺ uptake (left) and uptake rate (right) (Circle 2) ( k ) ( n = 10, 9, 9, 6, 9, 4, 5 cells per group). One-way ANOVA followed by Tukey’s or Dunnett’s ( k ) post-hoc multiple comparison test. All data are presented as mean ± S.E.M. Source data are provided as a file.

    Article Snippet: The following primary antibodies were used in this study: ACTB (Santa Cruz Biotechnology; sc-47778; lot # J0421; WB 1:5000), TUBA (Santa Cruz Biotechnology; sc-23948; lot # G2921; WB 1:5000), TUSC3 (clone D-9; Santa Cruz Biotechnology; sc-390566; lot # J1218; IF 1:500), GRP78 (Santa Cruz Biotechnology; sc-376768; lot # C0316; WB 1:3000), p-CREB-1-S133 (clone 10E9; Santa Cruz Biotechnology; sc-81486; WB 1:3000), FLAG (Sigma-Aldrich; F1804; lot # 0000375542; WB 1:3000), RFP (MBL; PM005; lot # 048; WB 1:3000), TUSC3 (Proteintech; 16039-1-AP; lot # 00007251; WB 1:500), AKT (Cell Signaling Technology; 9272; lot # 6; WB 1:3000), P-AKT (S473) (Cell Signaling Technology; 4060; lot # 27; WB 1:3000; IF 1:250), CREB (Cell Signaling Technology; 9197; WB 1:1000), CHOP (Cell Signaling Technology; 2895; WB 1:1000), ATF6 (Cell Signaling Technology; 65880; lot # 5; WB 1:3000), MAP2 (Cell Signaling Technology; 4542; IF 1:500), NCAM1 (Cell Signaling Technology; 3576; WB 1:3000), PERK (ABclonal; A18196; lot # 3560572205; WB 1:3000), p-PERK (T982) (ABclonal; AP0886; lot # 5500040024; WB 1:3000), XBP1s (ABclonal; A17007; lot # 1153240401; WB 1:3000), eIF2α (ABclonal; A21221; lot # 360002151; WB 1:3000), p-eIF2α (ABclonal; AP0745; lot # 360003834; WB 1:3000), Puromycin (ABclonal; A23931; lot # 3523033102; WB 1:3000), PSD-93 (ABclonal; A19669; WB 1:3000), PSD-95 (ABclonal; A7889; WB 1:3000), GluA1 (ABclonal; A1826; lot # 3561869003; WB 1:3000; IF 1:250), NMDAR1 (ABclonal; A7677; lot # 5500016744; WB 1:1000), Synaptophysin (ABclonal; A6344; lot # 5500008363; WB 1:1000; IF 1:250), and N-Cadherin (BD Biosciences; 610920; WB 1:3000).

    Techniques: Transfection, Confocal Microscopy, Immunoprecipitation, Comparison

    a Representative western blot of ER stress proteins in the hippocampus of 4-month-old female WT and TUSC3 KO mice. b – i Quantification of GRP78 ( b ), p-PERK (T982) ( c ), p-eIF2α (S51) ( d ), CHOP ( e ), XBP1s ( f ), p-AKT (S473) ( g ), p-CREB (S133) ( h ), and ATF6 ( i ) normalized to β-actin ( n = 3 mice per group). j Representative western blot analysis of ER stress proteins in the striatum of 4-month-old female WT and TUSC3 KO mice ( n = 3 per group). k – r Quantification of GRP78 ( k ), p-PERK (T982) ( l ), p-eIF2α (S51) ( m ), CHOP ( n ), XBP1s ( o ), p-AKT (S473) ( p ), p-CREB (S133) ( q ), and ATF6 ( r ) normalized to β-actin ( n = 3 mice per group). s Non-ID and ID1/ID2 fibroblasts were chronically treated with MgT (400 μM) for 48 h and analyzed by western blotting (left). Quantification of p-eIF2α (S51) levels normalized to eIF2α (right) ( n = 3 independent biological replicates). t Non-ID and ID-derived fibroblasts were pretreated with or without MgT (400 μM) for 24 h, followed by puromycin (5 μg/ml) treatment for 2 h. Representative western blot using an anti-puromycin antibody (left), and puromycin incorporation was quantified and normalized to β-actin (right) ( n = 3 independent biological replicates). u SH-SY5Y shControl and shTUSC3#3-1 cells were pretreated with MgT (400 μM) for 12 h, then treated with either DMSO, thapsigargin (2 μM), tunicamycin (2 μg/ml), A23187 (2 μM), or etoposide (25 μM) for 24 h. Cells were stained with propidium iodide and Calcein-AM, and double-positive cells were quantified and normalized to Calcein-AM-positive cells. Data represent three independent biological experiments ( n = 3). The exact number of cells analyzed per condition is provided in the Source Data file. v Representative western blot of primary cortical neurons (DIV7) treated with vehicle or 4-PBA (10 mM) for 24 h (left). Quantification of indicated protein levels normalized to TUBA (right) ( n = 3 independent biological replicates). Two-tailed unpaired t -test ( b – i , and k – r ); one-way ANOVA followed by Tukey’s post hoc multiple comparison test ( s , t , v ); two-way ANOVA followed by Tukey’s multiple comparison test ( u ). Data are presented as mean ± S.E.M. or S.D. ( t , u ). Source data are provided as a file.

    Journal: Nature Communications

    Article Title: TUSC3 regulates ERMA-mediated Mg 2+ uptake for synaptic function and neurodevelopment

    doi: 10.1038/s41467-025-65668-1

    Figure Lengend Snippet: a Representative western blot of ER stress proteins in the hippocampus of 4-month-old female WT and TUSC3 KO mice. b – i Quantification of GRP78 ( b ), p-PERK (T982) ( c ), p-eIF2α (S51) ( d ), CHOP ( e ), XBP1s ( f ), p-AKT (S473) ( g ), p-CREB (S133) ( h ), and ATF6 ( i ) normalized to β-actin ( n = 3 mice per group). j Representative western blot analysis of ER stress proteins in the striatum of 4-month-old female WT and TUSC3 KO mice ( n = 3 per group). k – r Quantification of GRP78 ( k ), p-PERK (T982) ( l ), p-eIF2α (S51) ( m ), CHOP ( n ), XBP1s ( o ), p-AKT (S473) ( p ), p-CREB (S133) ( q ), and ATF6 ( r ) normalized to β-actin ( n = 3 mice per group). s Non-ID and ID1/ID2 fibroblasts were chronically treated with MgT (400 μM) for 48 h and analyzed by western blotting (left). Quantification of p-eIF2α (S51) levels normalized to eIF2α (right) ( n = 3 independent biological replicates). t Non-ID and ID-derived fibroblasts were pretreated with or without MgT (400 μM) for 24 h, followed by puromycin (5 μg/ml) treatment for 2 h. Representative western blot using an anti-puromycin antibody (left), and puromycin incorporation was quantified and normalized to β-actin (right) ( n = 3 independent biological replicates). u SH-SY5Y shControl and shTUSC3#3-1 cells were pretreated with MgT (400 μM) for 12 h, then treated with either DMSO, thapsigargin (2 μM), tunicamycin (2 μg/ml), A23187 (2 μM), or etoposide (25 μM) for 24 h. Cells were stained with propidium iodide and Calcein-AM, and double-positive cells were quantified and normalized to Calcein-AM-positive cells. Data represent three independent biological experiments ( n = 3). The exact number of cells analyzed per condition is provided in the Source Data file. v Representative western blot of primary cortical neurons (DIV7) treated with vehicle or 4-PBA (10 mM) for 24 h (left). Quantification of indicated protein levels normalized to TUBA (right) ( n = 3 independent biological replicates). Two-tailed unpaired t -test ( b – i , and k – r ); one-way ANOVA followed by Tukey’s post hoc multiple comparison test ( s , t , v ); two-way ANOVA followed by Tukey’s multiple comparison test ( u ). Data are presented as mean ± S.E.M. or S.D. ( t , u ). Source data are provided as a file.

    Article Snippet: The following primary antibodies were used in this study: ACTB (Santa Cruz Biotechnology; sc-47778; lot # J0421; WB 1:5000), TUBA (Santa Cruz Biotechnology; sc-23948; lot # G2921; WB 1:5000), TUSC3 (clone D-9; Santa Cruz Biotechnology; sc-390566; lot # J1218; IF 1:500), GRP78 (Santa Cruz Biotechnology; sc-376768; lot # C0316; WB 1:3000), p-CREB-1-S133 (clone 10E9; Santa Cruz Biotechnology; sc-81486; WB 1:3000), FLAG (Sigma-Aldrich; F1804; lot # 0000375542; WB 1:3000), RFP (MBL; PM005; lot # 048; WB 1:3000), TUSC3 (Proteintech; 16039-1-AP; lot # 00007251; WB 1:500), AKT (Cell Signaling Technology; 9272; lot # 6; WB 1:3000), P-AKT (S473) (Cell Signaling Technology; 4060; lot # 27; WB 1:3000; IF 1:250), CREB (Cell Signaling Technology; 9197; WB 1:1000), CHOP (Cell Signaling Technology; 2895; WB 1:1000), ATF6 (Cell Signaling Technology; 65880; lot # 5; WB 1:3000), MAP2 (Cell Signaling Technology; 4542; IF 1:500), NCAM1 (Cell Signaling Technology; 3576; WB 1:3000), PERK (ABclonal; A18196; lot # 3560572205; WB 1:3000), p-PERK (T982) (ABclonal; AP0886; lot # 5500040024; WB 1:3000), XBP1s (ABclonal; A17007; lot # 1153240401; WB 1:3000), eIF2α (ABclonal; A21221; lot # 360002151; WB 1:3000), p-eIF2α (ABclonal; AP0745; lot # 360003834; WB 1:3000), Puromycin (ABclonal; A23931; lot # 3523033102; WB 1:3000), PSD-93 (ABclonal; A19669; WB 1:3000), PSD-95 (ABclonal; A7889; WB 1:3000), GluA1 (ABclonal; A1826; lot # 3561869003; WB 1:3000; IF 1:250), NMDAR1 (ABclonal; A7677; lot # 5500016744; WB 1:1000), Synaptophysin (ABclonal; A6344; lot # 5500008363; WB 1:1000; IF 1:250), and N-Cadherin (BD Biosciences; 610920; WB 1:3000).

    Techniques: Western Blot, Derivative Assay, Staining, Two Tailed Test, Comparison

    a , b Schematic of the Y-maze test ( a ) and spatial memory performance in WT and TUSC3 KO mice ( b ). Created with BioRender.com. Schematic of the novel object recognition (NOR) test ( c ) and representative tracking plots in the open field in the NOR test ( d ). Created with BioRender.com. e The NOR test (discrimination index) for WT and TUSC3 KO mice. f , g Schematic of the forced swim test (FST), consisting of a 120 s adaptation phase followed by 240 s of immobility measurement ( f ) and immobility time in the FST, assessing stress-coping ability in WT and TUSC3 KO mice ( g ). Created with BioRender.com. h , i Schematic of the tail suspension test (TST), with immobility recorded over 360 s ( h ) and immobility time in the TST for WT and TUSC3 KO mice ( i ). Created with BioRender.com. j , k Schematic of three-chambered social interaction test ( j ) and representative tracking plots from the social interaction test, showing times spent in each chamber ( k ). Created with BioRender.com. l , m Social interaction preference in TUSC3 KO mice, assessed by time spent with the novel mouse ( l ) and sniffing time ( m ). n – q Schematic of three-chambered social preference test ( n ) and representative tracking plots of WT and TUSC3 KO mice in social preference test ( o ). Social preference for the novel mouse of WT and TUSC3 KO mice in the social preference test ( p ). Sniffing time data confirms this lack of preference ( q ). Created with BioRender.com. In all behavioral experiments, 4-month-old mice were used: WT: 9 (5 F, 4 M); KO: 10 (6 F, 4 M). Two-tailed unpaired t -test ( b , e , g , i ), Two-way ANOVA followed by Tukey’s post-hoc multiple comparison test ( l , m , p , q ). All data are presented as mean ± S.E.M. Source data are provided as a file.

    Journal: Nature Communications

    Article Title: TUSC3 regulates ERMA-mediated Mg 2+ uptake for synaptic function and neurodevelopment

    doi: 10.1038/s41467-025-65668-1

    Figure Lengend Snippet: a , b Schematic of the Y-maze test ( a ) and spatial memory performance in WT and TUSC3 KO mice ( b ). Created with BioRender.com. Schematic of the novel object recognition (NOR) test ( c ) and representative tracking plots in the open field in the NOR test ( d ). Created with BioRender.com. e The NOR test (discrimination index) for WT and TUSC3 KO mice. f , g Schematic of the forced swim test (FST), consisting of a 120 s adaptation phase followed by 240 s of immobility measurement ( f ) and immobility time in the FST, assessing stress-coping ability in WT and TUSC3 KO mice ( g ). Created with BioRender.com. h , i Schematic of the tail suspension test (TST), with immobility recorded over 360 s ( h ) and immobility time in the TST for WT and TUSC3 KO mice ( i ). Created with BioRender.com. j , k Schematic of three-chambered social interaction test ( j ) and representative tracking plots from the social interaction test, showing times spent in each chamber ( k ). Created with BioRender.com. l , m Social interaction preference in TUSC3 KO mice, assessed by time spent with the novel mouse ( l ) and sniffing time ( m ). n – q Schematic of three-chambered social preference test ( n ) and representative tracking plots of WT and TUSC3 KO mice in social preference test ( o ). Social preference for the novel mouse of WT and TUSC3 KO mice in the social preference test ( p ). Sniffing time data confirms this lack of preference ( q ). Created with BioRender.com. In all behavioral experiments, 4-month-old mice were used: WT: 9 (5 F, 4 M); KO: 10 (6 F, 4 M). Two-tailed unpaired t -test ( b , e , g , i ), Two-way ANOVA followed by Tukey’s post-hoc multiple comparison test ( l , m , p , q ). All data are presented as mean ± S.E.M. Source data are provided as a file.

    Article Snippet: The following primary antibodies were used in this study: ACTB (Santa Cruz Biotechnology; sc-47778; lot # J0421; WB 1:5000), TUBA (Santa Cruz Biotechnology; sc-23948; lot # G2921; WB 1:5000), TUSC3 (clone D-9; Santa Cruz Biotechnology; sc-390566; lot # J1218; IF 1:500), GRP78 (Santa Cruz Biotechnology; sc-376768; lot # C0316; WB 1:3000), p-CREB-1-S133 (clone 10E9; Santa Cruz Biotechnology; sc-81486; WB 1:3000), FLAG (Sigma-Aldrich; F1804; lot # 0000375542; WB 1:3000), RFP (MBL; PM005; lot # 048; WB 1:3000), TUSC3 (Proteintech; 16039-1-AP; lot # 00007251; WB 1:500), AKT (Cell Signaling Technology; 9272; lot # 6; WB 1:3000), P-AKT (S473) (Cell Signaling Technology; 4060; lot # 27; WB 1:3000; IF 1:250), CREB (Cell Signaling Technology; 9197; WB 1:1000), CHOP (Cell Signaling Technology; 2895; WB 1:1000), ATF6 (Cell Signaling Technology; 65880; lot # 5; WB 1:3000), MAP2 (Cell Signaling Technology; 4542; IF 1:500), NCAM1 (Cell Signaling Technology; 3576; WB 1:3000), PERK (ABclonal; A18196; lot # 3560572205; WB 1:3000), p-PERK (T982) (ABclonal; AP0886; lot # 5500040024; WB 1:3000), XBP1s (ABclonal; A17007; lot # 1153240401; WB 1:3000), eIF2α (ABclonal; A21221; lot # 360002151; WB 1:3000), p-eIF2α (ABclonal; AP0745; lot # 360003834; WB 1:3000), Puromycin (ABclonal; A23931; lot # 3523033102; WB 1:3000), PSD-93 (ABclonal; A19669; WB 1:3000), PSD-95 (ABclonal; A7889; WB 1:3000), GluA1 (ABclonal; A1826; lot # 3561869003; WB 1:3000; IF 1:250), NMDAR1 (ABclonal; A7677; lot # 5500016744; WB 1:1000), Synaptophysin (ABclonal; A6344; lot # 5500008363; WB 1:1000; IF 1:250), and N-Cadherin (BD Biosciences; 610920; WB 1:3000).

    Techniques: Suspension, Two Tailed Test, Comparison

    a Representative western blot of PSD-93, PSD-95, GluA1, and phosphorylated CREB (S133) in whole-brain lysates of 4-month-old WT and TUSC3 KO female mice (left). Quantification of indicated protein levels (right) ( n = 3). b , c Representative confocal images of immunohistochemistry for TUSC3, synaptophysin (SYP), and MAP2 in the CA1 and CA3 hippocampal regions and the striatum of 4-month-old WT and TUSC3 KO female mice ( b ), and quantification of SYP intensity normalized to MAP2 ( c ) ( n = 3). Scale bar: 50 μm. d , e Representative confocal images of immunohistochemistry for TUSC3, GluA1, and MAP2 in the CA1 and CA3 hippocampal regions and the striatum of 4-month-old WT and TUSC3 KO female mice ( d ), and quantification of GluA1 intensity normalized to MAP2 ( e ) ( n = 3). Scale bar: 50 μm. f , g Representative dendritic images of primary hippocampal neurons transfected with EGFP ( f ). Scale bar: 50 μm. Sholl analysis of dendritic branching in WT and TUSC3 KO neurons ( n = 20 cells per group) ( g ). h , i Representative confocal images of dendritic spines of primary hippocampal neurons (DIV 12) transfected with EGFP for 48 h ( h ). Violin plot quantifying dendritic spine density (spines per 10 μm) in WT and KO neurons ( n = 20 cells per group) ( i ). Scale bar: 10 μm. j–l Representative traces of sEPSCs recorded from hippocampal slices of 2-month-old WT and TUSC3 KO mice ( j ), quantification of sEPSC amplitude ( k ), and frequency ( l ) in WT ( n = 12 cells from 5 mice; 4 F, 1 M) and TUSC3 KO ( n = 8 cells from 5 mice; 2 F, 3 M). m Representative traces (left) and quantification of the eEPSC amplitude (right) in WT ( n = 8 cells from 5 mice; 4 F, 1 M), TUSC3 KO ( n = 8 cells from 5 mice; 2 F, 3 M). n Representative traces (left) and EPSC-PPR (right) in WT ( n = 8 cells from 5 mice; 3 F, 2 M), TUSC3 KO ( n = 9 cells from 6 mice; 4 F, 2 M). Two-tailed unpaired t -test. Data are presented as mean ± S.E.M. ( a , c , e , and k – n ) or S.D. ( g , i ). Source data are provided as a file.

    Journal: Nature Communications

    Article Title: TUSC3 regulates ERMA-mediated Mg 2+ uptake for synaptic function and neurodevelopment

    doi: 10.1038/s41467-025-65668-1

    Figure Lengend Snippet: a Representative western blot of PSD-93, PSD-95, GluA1, and phosphorylated CREB (S133) in whole-brain lysates of 4-month-old WT and TUSC3 KO female mice (left). Quantification of indicated protein levels (right) ( n = 3). b , c Representative confocal images of immunohistochemistry for TUSC3, synaptophysin (SYP), and MAP2 in the CA1 and CA3 hippocampal regions and the striatum of 4-month-old WT and TUSC3 KO female mice ( b ), and quantification of SYP intensity normalized to MAP2 ( c ) ( n = 3). Scale bar: 50 μm. d , e Representative confocal images of immunohistochemistry for TUSC3, GluA1, and MAP2 in the CA1 and CA3 hippocampal regions and the striatum of 4-month-old WT and TUSC3 KO female mice ( d ), and quantification of GluA1 intensity normalized to MAP2 ( e ) ( n = 3). Scale bar: 50 μm. f , g Representative dendritic images of primary hippocampal neurons transfected with EGFP ( f ). Scale bar: 50 μm. Sholl analysis of dendritic branching in WT and TUSC3 KO neurons ( n = 20 cells per group) ( g ). h , i Representative confocal images of dendritic spines of primary hippocampal neurons (DIV 12) transfected with EGFP for 48 h ( h ). Violin plot quantifying dendritic spine density (spines per 10 μm) in WT and KO neurons ( n = 20 cells per group) ( i ). Scale bar: 10 μm. j–l Representative traces of sEPSCs recorded from hippocampal slices of 2-month-old WT and TUSC3 KO mice ( j ), quantification of sEPSC amplitude ( k ), and frequency ( l ) in WT ( n = 12 cells from 5 mice; 4 F, 1 M) and TUSC3 KO ( n = 8 cells from 5 mice; 2 F, 3 M). m Representative traces (left) and quantification of the eEPSC amplitude (right) in WT ( n = 8 cells from 5 mice; 4 F, 1 M), TUSC3 KO ( n = 8 cells from 5 mice; 2 F, 3 M). n Representative traces (left) and EPSC-PPR (right) in WT ( n = 8 cells from 5 mice; 3 F, 2 M), TUSC3 KO ( n = 9 cells from 6 mice; 4 F, 2 M). Two-tailed unpaired t -test. Data are presented as mean ± S.E.M. ( a , c , e , and k – n ) or S.D. ( g , i ). Source data are provided as a file.

    Article Snippet: The following primary antibodies were used in this study: ACTB (Santa Cruz Biotechnology; sc-47778; lot # J0421; WB 1:5000), TUBA (Santa Cruz Biotechnology; sc-23948; lot # G2921; WB 1:5000), TUSC3 (clone D-9; Santa Cruz Biotechnology; sc-390566; lot # J1218; IF 1:500), GRP78 (Santa Cruz Biotechnology; sc-376768; lot # C0316; WB 1:3000), p-CREB-1-S133 (clone 10E9; Santa Cruz Biotechnology; sc-81486; WB 1:3000), FLAG (Sigma-Aldrich; F1804; lot # 0000375542; WB 1:3000), RFP (MBL; PM005; lot # 048; WB 1:3000), TUSC3 (Proteintech; 16039-1-AP; lot # 00007251; WB 1:500), AKT (Cell Signaling Technology; 9272; lot # 6; WB 1:3000), P-AKT (S473) (Cell Signaling Technology; 4060; lot # 27; WB 1:3000; IF 1:250), CREB (Cell Signaling Technology; 9197; WB 1:1000), CHOP (Cell Signaling Technology; 2895; WB 1:1000), ATF6 (Cell Signaling Technology; 65880; lot # 5; WB 1:3000), MAP2 (Cell Signaling Technology; 4542; IF 1:500), NCAM1 (Cell Signaling Technology; 3576; WB 1:3000), PERK (ABclonal; A18196; lot # 3560572205; WB 1:3000), p-PERK (T982) (ABclonal; AP0886; lot # 5500040024; WB 1:3000), XBP1s (ABclonal; A17007; lot # 1153240401; WB 1:3000), eIF2α (ABclonal; A21221; lot # 360002151; WB 1:3000), p-eIF2α (ABclonal; AP0745; lot # 360003834; WB 1:3000), Puromycin (ABclonal; A23931; lot # 3523033102; WB 1:3000), PSD-93 (ABclonal; A19669; WB 1:3000), PSD-95 (ABclonal; A7889; WB 1:3000), GluA1 (ABclonal; A1826; lot # 3561869003; WB 1:3000; IF 1:250), NMDAR1 (ABclonal; A7677; lot # 5500016744; WB 1:1000), Synaptophysin (ABclonal; A6344; lot # 5500008363; WB 1:1000; IF 1:250), and N-Cadherin (BD Biosciences; 610920; WB 1:3000).

    Techniques: Western Blot, Immunohistochemistry, Transfection, Two Tailed Test

    a Experimental timeline. Four-week-old mice received chronic magnesium supplementation via drinking water containing MgT (910 mg/kg/day) for 42 days, followed by behavioral analysis. Created with BioRender.com. b , c Cognitive performance of MgT-treated WT and TUSC3 KO mice assessed in the Y-maze ( b ) and NOR test ( c ). d Stress-coping ability evaluated using the TST. e , f Social behavior assessed in the social interaction ( e ) and social novelty preference ( f ) tests (left). Time spent in each chamber (middle) and sniffing time (right) were measured. g , h Representative western blot of hippocampal tissues from WT and TUSC3 KO mice treated with vehicle or MgT ( g ). Quantification of indicated protein levels ( h ) ( n = 3). i , j Representative confocal images of GluA1 immunohistochemistry in the CA1 ( i ) and CA3 ( j ) hippocampal regions from WT and TUSC3 KO mice treated with vehicle or MgT ( n = 3). Scale bar: 50 μm. In these experiments, 10-week-old mice were used: WT: 8 (4 F, 4 M); WT + MgT: 8 (4 F, 4 M); KO: 6 (3 F, 3 M); KO + MgT: 8 (5 F, 3 M). Two-way ANOVA with Tukey’s post hoc multiple comparison test. All data are represented as mean ± S.E.M. Source data are provided as a file.

    Journal: Nature Communications

    Article Title: TUSC3 regulates ERMA-mediated Mg 2+ uptake for synaptic function and neurodevelopment

    doi: 10.1038/s41467-025-65668-1

    Figure Lengend Snippet: a Experimental timeline. Four-week-old mice received chronic magnesium supplementation via drinking water containing MgT (910 mg/kg/day) for 42 days, followed by behavioral analysis. Created with BioRender.com. b , c Cognitive performance of MgT-treated WT and TUSC3 KO mice assessed in the Y-maze ( b ) and NOR test ( c ). d Stress-coping ability evaluated using the TST. e , f Social behavior assessed in the social interaction ( e ) and social novelty preference ( f ) tests (left). Time spent in each chamber (middle) and sniffing time (right) were measured. g , h Representative western blot of hippocampal tissues from WT and TUSC3 KO mice treated with vehicle or MgT ( g ). Quantification of indicated protein levels ( h ) ( n = 3). i , j Representative confocal images of GluA1 immunohistochemistry in the CA1 ( i ) and CA3 ( j ) hippocampal regions from WT and TUSC3 KO mice treated with vehicle or MgT ( n = 3). Scale bar: 50 μm. In these experiments, 10-week-old mice were used: WT: 8 (4 F, 4 M); WT + MgT: 8 (4 F, 4 M); KO: 6 (3 F, 3 M); KO + MgT: 8 (5 F, 3 M). Two-way ANOVA with Tukey’s post hoc multiple comparison test. All data are represented as mean ± S.E.M. Source data are provided as a file.

    Article Snippet: The following primary antibodies were used in this study: ACTB (Santa Cruz Biotechnology; sc-47778; lot # J0421; WB 1:5000), TUBA (Santa Cruz Biotechnology; sc-23948; lot # G2921; WB 1:5000), TUSC3 (clone D-9; Santa Cruz Biotechnology; sc-390566; lot # J1218; IF 1:500), GRP78 (Santa Cruz Biotechnology; sc-376768; lot # C0316; WB 1:3000), p-CREB-1-S133 (clone 10E9; Santa Cruz Biotechnology; sc-81486; WB 1:3000), FLAG (Sigma-Aldrich; F1804; lot # 0000375542; WB 1:3000), RFP (MBL; PM005; lot # 048; WB 1:3000), TUSC3 (Proteintech; 16039-1-AP; lot # 00007251; WB 1:500), AKT (Cell Signaling Technology; 9272; lot # 6; WB 1:3000), P-AKT (S473) (Cell Signaling Technology; 4060; lot # 27; WB 1:3000; IF 1:250), CREB (Cell Signaling Technology; 9197; WB 1:1000), CHOP (Cell Signaling Technology; 2895; WB 1:1000), ATF6 (Cell Signaling Technology; 65880; lot # 5; WB 1:3000), MAP2 (Cell Signaling Technology; 4542; IF 1:500), NCAM1 (Cell Signaling Technology; 3576; WB 1:3000), PERK (ABclonal; A18196; lot # 3560572205; WB 1:3000), p-PERK (T982) (ABclonal; AP0886; lot # 5500040024; WB 1:3000), XBP1s (ABclonal; A17007; lot # 1153240401; WB 1:3000), eIF2α (ABclonal; A21221; lot # 360002151; WB 1:3000), p-eIF2α (ABclonal; AP0745; lot # 360003834; WB 1:3000), Puromycin (ABclonal; A23931; lot # 3523033102; WB 1:3000), PSD-93 (ABclonal; A19669; WB 1:3000), PSD-95 (ABclonal; A7889; WB 1:3000), GluA1 (ABclonal; A1826; lot # 3561869003; WB 1:3000; IF 1:250), NMDAR1 (ABclonal; A7677; lot # 5500016744; WB 1:1000), Synaptophysin (ABclonal; A6344; lot # 5500008363; WB 1:1000; IF 1:250), and N-Cadherin (BD Biosciences; 610920; WB 1:3000).

    Techniques: Western Blot, Immunohistochemistry, Comparison

    TUSC3 deficiency impairs ER magnesium uptake by disrupting its positive modulation of ERMA, an ER Mg²⁺ transporter. This leads to reduced ER [Mg²⁺], activation of ER stress via the PERK–eIF2α axis, and suppression of global translation, including synaptic proteins (PSD-93, PSD-95, GluA1). In parallel, CREB activity is reduced through impaired PI3K/AKT signaling. Together, these changes result in synaptic dysfunction, increased neuronal vulnerability (via CHOP), and ultimately cognitive and adaptive behavioral deficits. Notably, magnesium supplementation restores ER [Mg²⁺] levels, alleviates ER stress, and rescues synaptic and cognitive phenotypes in TUSC3-deficient mice, suggesting a potential therapeutic strategy for TUSC3-associated ID.

    Journal: Nature Communications

    Article Title: TUSC3 regulates ERMA-mediated Mg 2+ uptake for synaptic function and neurodevelopment

    doi: 10.1038/s41467-025-65668-1

    Figure Lengend Snippet: TUSC3 deficiency impairs ER magnesium uptake by disrupting its positive modulation of ERMA, an ER Mg²⁺ transporter. This leads to reduced ER [Mg²⁺], activation of ER stress via the PERK–eIF2α axis, and suppression of global translation, including synaptic proteins (PSD-93, PSD-95, GluA1). In parallel, CREB activity is reduced through impaired PI3K/AKT signaling. Together, these changes result in synaptic dysfunction, increased neuronal vulnerability (via CHOP), and ultimately cognitive and adaptive behavioral deficits. Notably, magnesium supplementation restores ER [Mg²⁺] levels, alleviates ER stress, and rescues synaptic and cognitive phenotypes in TUSC3-deficient mice, suggesting a potential therapeutic strategy for TUSC3-associated ID.

    Article Snippet: The following primary antibodies were used in this study: ACTB (Santa Cruz Biotechnology; sc-47778; lot # J0421; WB 1:5000), TUBA (Santa Cruz Biotechnology; sc-23948; lot # G2921; WB 1:5000), TUSC3 (clone D-9; Santa Cruz Biotechnology; sc-390566; lot # J1218; IF 1:500), GRP78 (Santa Cruz Biotechnology; sc-376768; lot # C0316; WB 1:3000), p-CREB-1-S133 (clone 10E9; Santa Cruz Biotechnology; sc-81486; WB 1:3000), FLAG (Sigma-Aldrich; F1804; lot # 0000375542; WB 1:3000), RFP (MBL; PM005; lot # 048; WB 1:3000), TUSC3 (Proteintech; 16039-1-AP; lot # 00007251; WB 1:500), AKT (Cell Signaling Technology; 9272; lot # 6; WB 1:3000), P-AKT (S473) (Cell Signaling Technology; 4060; lot # 27; WB 1:3000; IF 1:250), CREB (Cell Signaling Technology; 9197; WB 1:1000), CHOP (Cell Signaling Technology; 2895; WB 1:1000), ATF6 (Cell Signaling Technology; 65880; lot # 5; WB 1:3000), MAP2 (Cell Signaling Technology; 4542; IF 1:500), NCAM1 (Cell Signaling Technology; 3576; WB 1:3000), PERK (ABclonal; A18196; lot # 3560572205; WB 1:3000), p-PERK (T982) (ABclonal; AP0886; lot # 5500040024; WB 1:3000), XBP1s (ABclonal; A17007; lot # 1153240401; WB 1:3000), eIF2α (ABclonal; A21221; lot # 360002151; WB 1:3000), p-eIF2α (ABclonal; AP0745; lot # 360003834; WB 1:3000), Puromycin (ABclonal; A23931; lot # 3523033102; WB 1:3000), PSD-93 (ABclonal; A19669; WB 1:3000), PSD-95 (ABclonal; A7889; WB 1:3000), GluA1 (ABclonal; A1826; lot # 3561869003; WB 1:3000; IF 1:250), NMDAR1 (ABclonal; A7677; lot # 5500016744; WB 1:1000), Synaptophysin (ABclonal; A6344; lot # 5500008363; WB 1:1000; IF 1:250), and N-Cadherin (BD Biosciences; 610920; WB 1:3000).

    Techniques: Activation Assay, Activity Assay