Fig. 2: TUSC3 regulates ER Mg²⁺ uptake by interacting with ERMA.
From: TUSC3 regulates ERMA-mediated Mg2+ uptake for synaptic function and neurodevelopment
a–d SH-SY5Y shControl and shTUSC3#3-1 cells were co-transfected with Mag-FRETER 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-FRETER 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 Source data file.
