Supplementary Figure 1: CCCP and L.m induce mitophagy response in mouse PMs.
From: Listeria hijacks host mitophagy through a novel mitophagy receptor to evade killing
a,b, Q-PCR analysis of mtDNA/nDNA (a) (n=3) and western blot analysis of mitochondria markers TIM23 and HSP60 (b) in mouse PMs after CCCP stimulation (30μM) for the indicated time points. c,d, Mitochondria membrane potential (Δψm) determined by TMRM staining after L.m infection (c) or CCCP (30μM) stimulation (d). (n=3). e, Q-PCR analysis of mtDNA/nDNA in mouse PMs infected with L.m at MOI 5 with or without CQ (n=3). f, Western blot analysis of mitochondria markers TIM23 and HSP60 treated as in (e). g, Light microscopy analysis of mouse PM morphology after L.m infection at the indicated time points at MOI 5. TNF+ZVAD as a positive control for cell death. Scale bars, 100μm. h,i, Western blot analysis of cleaved caspase-3 (h) or caspase-1 (i) after L.m infection at the indicated time points at MOI 5 in PMs. TNF+CHX (TC) and LPS+ATP (L+A) as positive controls. j. LDH release from mouse PMs after L.m infection at MOI 5 for the indicated time points or CCCP treatment (n=4). TNF+ZVAD (TZ) as a positive control. k, Q-PCR analysis of mtDNA/nDNA of mouse PMs infected with L.m and treated with or without Mdivi-1 (20μM) for 3h (n=3). l, Δψm after L.m infection or CCCP stimulation for 6h in the THP1 macrophages (n=3). m, Q-PCR analysis of mtDNA/nDNA in THP1 macrophages infected with L.m at MOI 5 with or without CQ (n=3). n, Western blot analysis of mitochondria markers TIM23 and HSP60 treated as in (m). o, Q-PCR analysis of mtDNA/nDNA of THP1 macrophages infected with L.m and treated with or without Mdivi-1 for 3h (n=3). Data are representative of three (a-l) or two independent experiments (m-o) and represent mean ± SEM (a,c-e,j-m,o). Two-sided Student’s t-test was used to measure significance.
