Journal: bioRxiv

Article Title: Chronic cold exposure induces plasticity of mitochondrial calcium uptake in beige and brown fat of UCP1-deficient mice

doi: 10.64898/2026.03.16.712209

Figure Lengend Snippet: A. Swelling of BAT mitochondria from WT and Ucp1 -/- mice upon 3 weeks of cold exposure. B. Swelling of mitochondria from liver of WT mice kept at RT for comparison. C. Immunoblots of proteins implicated in ERMCs: GRP75, IP3R, VDAC, and cyclophilin D (CyD). D. Quantified protein expression. Calnexin, TOM20 and Na/K ATPase were used for normalization per ER biomass, mitochondrial biomass, and total extract, respectively. E. Representative image of PLA for IP3R–VDAC in BAT of WT and Ucp1 -/- mice exposed to 6°C for 3 weeks. F. Quantification of the contact sites from E . n = 4-6 mice. Data shown as mean ± sem. Unpaired t-test.

Article Snippet: Lysates were resolved by sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis (PAGE), transferred to a polyvinylidene difluoride (PVDF) membrane (Millipore), and probed with antibodies against Na + /K + -ATPase (Abcam, ab76020, GR3237646-13, 1/10,000), TOMM20 (Sigma Prestige, HPA011562, 1/500), MCU (Cell Signaling Technology, 14997, 1/100), UCP1 (Abcam, ab10983, 1/1000), NCLX (Proteintech, 21430-1-AP, 1/500), TMBIM5 (Proteintech, 16296-1-AP, 1/500), TMEM65 (Invitrogen, PA5-112762, 1/500), GRP75 (Abcam, ab53098, 1/500), Calnexin (BD Transduction Laboratories, 610523, 1/1000), VDAC (Cell Signaling Technology, 4661, 1/2000), CyD (Proteintech, 18466-1-AP, 1/1000), OXPHOS (Abcam, ab110413, 1/500), Vinculin (1:10000; V9131; Sigma-Aldrich), NDUFB6 (1:5000; #6510832 Thermo Fisher Scientific), mtCOI (1:10000; #10444035; Thermo Fisher Scientific), ATP5A (1:1,000; #439800; Thermo Fisher Scientific), SDHA (1:1,000, #459200, Thermo Fisher Scientific), SDHB (1:1,000, #ab14714, Abcam), UQCRC2 (1:1,000, #14742-1-AP; Proteintech), ATPIF1 (1:1,000, #13268S; Cell Signaling Technology), TOMM20 Alexa Fluor 488 (1:1,000; #ab205486; Abcam), and cytochrome c (1:5,000, #ab110325, Abcam).

Techniques: Comparison, Western Blot, Expressing