Fig. 3: Immune cells of mice surviving sepsis exhibit alterations in mitochondrial and cellular Ca²⁺ regulation.

C57Bl/6 mice underwent Sham or CLP and best clinical practices. A Macrophage (Mφ) MCU complex expression. At day 30, peritoneal Mφ were isolated, and total cell protein lysate was analyzed by immunoblot for MICU1, MICU2, MCUR1, MCU, TOM20, and actin. Sham (n = 2 mice). CLP (n = 5 mice) B Mφ mitochondrial [Ca²⁺]_m ex vivo. At 30 days, peritoneal Mφ were isolated from Sham and CLP mice, plated, and then loaded with Rhod-2 am for 30 min at 37 °C. Fluorescence intensity as a marker of [Ca²⁺]_m was imaged by spectrophotometry: ex/em 552 nm/581 nm. Sham (n = 4 mice) CLP (n = 5 mice) C siRNA inhibition of MICU1 and MCUR1. RAW 264.7 Mφ cells were transfected with Non-target (NT), MICU1, MCUR1, or both MICU1 and MCUR1 siRNA. After 72 h, cells were loaded with Rhod-2 am at 37 °C. Fluorescence intensity was imaged by spectrophotometry: ex/em 552 nm/581 nm. In parallel cell populations, total cell protein lysate was isolated and analyzed by immunoblot for MICU1-3, MCUR1, MCU, TOM20, and actin. RAW 264.7 (4 separate experiments; n = 3 and 4 cell groups per experiment) Wilcoxon rank sum. bar = median.