ZFIN Image: Pagán et al., 2022, Figure S2

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Figure Caption

Figure S2

Inhibition of autophagic cell death, mitochondrial apoptosis, or TNF-associated necrosis does not prevent mycobacterium-induced macrophage death in mTOR-deficient animals, related to Figures 2 and 3

(A) Confocal micrographs of LC3 aggregation in neuromasts, clusters of mechanosensory cells of the fish lateral line, from atg12^sa42684 incross fish expressing Tg(CMV:lc3b-GFP) 5 dpf. GFP fluorescence (top) and surface-rendered puncta (bottom) are shown. Scale bar, 10 μm.

(B) Number of LC3 puncta per neuromast.

(C) Cording in rapamycin- or DMSO-treated atg12^sa42684 incross fish 5 dpi.

(D) Confocal micrographs of acridine orange (AO) staining (green) and surface-rendered puncta (magenta) in the midbrain of casp9^sa11164 incross fish 3 dpf. Scale bars, 50 μm.

(E) Number of AO puncta in the midbrain.

(F) Cording in rapamycin- or DMSO-treated casp9^sa11164 incross fish 5 dpi.

(G) Cording in rapamycin- and DMSO-treated pycard^w216/w216 (Asc-deficient) animals and siblings 4 dpi.

(H) Cording in rapamycin- and DMSO-treated sting1^sa35634 (Sting-deficient) animals and WT siblings 5 dpi.

(I) Necrosis pathway induced by mycobacterial infection plus excess TNF and pharmacological interventions tested.

(J and K) Mycobacterial cording in mtor^fh178/+ incross fish treated with (H) necrostatin-1, (I) nifedipine (5 μM), diltiazem (10 μM), or 0.5% DMSO 4 dpi.

Symbols represent individual (B) neuromasts or (E) animals. (B and E) Horizontal lines indicate mean values. (C, F–H, J, and K) Numbers within columns indicate animals per group. (B and E) One-way ANOVA with Tukey’s post-test.

Acknowledgments

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Reprinted from Cell, 185(20), Pagán, A.J., Lee, L.J., Edwards-Hicks, J., Moens, C.B., Tobin, D.M., Busch-Nentwich, E.M., Pearce, E.L., Ramakrishnan, L., mTOR-regulated mitochondrial metabolism limits mycobacterium-induced cytotoxicity, 3720-3738.e13, Copyright (2022) with permission from Elsevier. Full text @ Cell