Figure 3.

CNNM4 mediates Mg release from thermogenic adipocytes. A) Representative staining of TMRM, MitoTracker, and Mag‐Fluo‐4 in beige adipocytes treated with indicated compounds (CL316243 2 µg mL⁻¹, H89 10 × 10⁻⁶ m, FCCP 5 × 10⁻⁶ m, CsA 100 × 10⁻⁹ m). Scale bar, 20 µm; B) Mg transmembrane transporter expression in adipocytes from mice under cold exposure or RT through single cell sequencing from literature; C) fold change of the Mg transporters gene expression (Mrs2, Cnnm4, Cnnm3, Slc41a1, Slc41a2, Magt1, Trpm6, and Trpm7) in scWAT, SVF, or adipocytes under cold exposure or CL316243 stimulation; D) UCP1 and CNNM4 protein expression in scWAT from mice under room temperature (25 °C) or cold exposure (4 °C); E–G) UCP1 and CNNM4 protein expression in scWAT from mice with 6‐OHDA injection, surgical transection of sympathetic neuron fibers, or local injection with L748337 under cold exposure; H) UCP1 and CNNM4 protein expression and CREB phosphorylation in primary beige adipocytes treated with CL316243 (2 µg mL⁻¹) together with L748337 (4 × 10⁻⁶ m) or H89 (10 × 10⁻⁶ m); I) ChIP‐qPCR analysis of CREB binding to CNNM4 promoter in primary beige adipocytes treated with CL316243. Result was normalized to input control values and represented as fold enrichment relative to the anti‐rabbit IgG control (n = 3); J) Mg levels in scWAT‐derived interstitial fluid from CNNM4^Adipoq‐cKO or WT mice (n = 6); K) Mg levels in scWAT‐derived interstitial fluid from C57BL/6 mice with local injection of Adv‐GFP or Adv‐CNNM4 (n = 6). Data were expressed as means ± SEM. (B), (C), (J), and (K) were calculated by unpaired two‐tailed Student's t‐test; (I) were calculated by two‐way ANOVA followed with Bonferroni's multiple comparison test.