Fig. 1: Functional characterisation and overall architecture of hSLC30A10.

a Normalized Mn levels in SH-SY5Y cells (n = 5). b, c Flow cytometry detection (b) and statistical analysis (c; n = 3) of cell survival by Annexin V / PI. d Cell survival by Annexin V / PI in the absence or the presence of 100 μM MnCl₂ for 24 h in Dox-inducible SLC30A10 overexpressed SH-SY5Y cells (n = 4). -Dox, without Dox treatment. +Dox, treated with 100 ng/ml Dox. e Normalized Mn levels in SLC30A10 transfected HEK293T cells by ICP-MS. Cells transfected with vector served as control. Data were normalized to control (n = 4). f In vitro Mn²⁺ transport assay of SLC30A10. Purified human SLC30A10 protein and calcein-salt were reconstituted into liposome. The fluorescence of calcein was quenched by Mn²⁺ that is transported into the liposome (n = 3). Transport was initiated by the addition of 100 μM MnCl₂. The fluorescence of MnCl₂-free proteoliposome was measured to determine the background signal (F₀). The fluorescence of 0 s was measured as maximum (max) fluorescence. The relative fluorescence was calculated using the equation (F/Fmax)-(1-F₀/F₀max). g, h Cryo-EM map of human SLC30A10 in the Mn²⁺-bound state in the side view. The two monomers are coloured in red and blue. i, j Cylinder representation of SLC30A10 in the Mn²⁺-bound state in the side view. Mn²⁺ is in purple spheres. The flexible region in the loops between TM4 and TM5 not observed in the structure are shown as dash line. k Cylinder representation of SLC30A10 in the Mn²⁺-bound state in the extracellular view. l Topology of SLC30A10 monomer. All bar graphs above represent mean ± SEM. Statistical analysis was performed using two-tailed unpaired t-test (a, c, e), one-way ANOVA with Dunnett’s multiple comparisons test (d), or two-way ANOVA with Sidak’s multiple comparisons test (f) (****p < 0.0001; ***p < 0.001; **p < 0.01; *p < 0.05; ns not significant). The experiments were independently repeated 3 times.