Extended Data Fig. 1: MCU Is required for L. monocytogenes-induced mitochondrial Ca²⁺ uptake in macrophages.

a−d, THP-1 cells (a and b) or BMMs (c and d) were incubated with rhod-2 (a and c) or Fluo-4 (b and d), followed by either WT or Δhly L. monocytogenes (MOI, 10) challenge. Fluorescence signal was read using a microplate reader. e−f, Fluorescence signal-based Ca²⁺ measurement to monitor mtCa²⁺ (e) and cytosolic Ca²⁺ (f) in Mcu ^fl/fl and Mcu ^∆mye BMMs upon L. monocytogenes challenge. g-h, mtCa²⁺ (g) and cytosolic Ca²⁺ (h) in Mcu ^fl/fl or Mcu ^∆mye BMMs were measured upon ATP stimulation (100 μM). i-j, mtCa²⁺ (i) and cytosolic Ca²⁺ (j) in Mcu ^fl/fl or Mcu ^∆mye BMMs were measured upon purified protein listeriolysin O (LLO) (0.5 nM) challenge. k, mtCa²⁺ in BMMs pretreated with Xestospongin C (5 μM) contained in normal DMEM or Ca²⁺ free DMEM for 2 hours, were measured upon 0.5 nM LLO protein challenge. l, mtCa²⁺ in BMMs pretreated with cytochalasin D (10 μM) were measured upon L. monocytogenes challenge. The results presented are presentative of three independent experiments.