To understand at the single-cell level whether the Legionella-induced ‘reverse mode’ of the mitochondrial FOF1-ATPase aids to conserve the Δψm during infection to delay cell death of infected macrophages, we simultaneously monitored Δψm and early cell death signs in the same infected cell by multiplexing Annexin-V and TMRM signals in our living cell assay (Figure 4E, Figure 4—figure supplement 2A and B). This evidence concerns the gene ANXA5 and infection.