asc is expressed during zebrafish early development. asc whole-mount, in situ hybridization (wish) of 3-dpf zebrafish larvae (A) with cross (B) and longitudinal (C) sectioning of plastic embedded wish sample showing expression in epidermis, intestinal epithelium, and cells located in the brain. Bars: (full larvae) 300 µm; (sections) 100 µm. Immunostaining of ASC in 3-dpf Tg(krt4:GFP) larva (D). Optical cross section of lateral fin showing GFP expression in the EVL and ASC expression on both epidermal layers (D′). WT 3-dpf larva immunostained for ASC, together with nuclear envelope marker lamin and DAPI shows its nuclear and cytoplasmic localization (E). Immunostaining of 3-dpf Tg(mpeg1:EGFP) (F), Tg(lyz:DsRed2) (G), and Tg(spi1b:GAL4,UAS:TagRFP) (H) larvae showing expression of ASC in macrophages, neutrophils, and a single myeloid cell in the caudal hematopoietic tissue (H, white arrowhead). Bars: (full larvae) 300 µm; (all others) 30 µm.
asc is expressed during zebrafish early development. asc whole-mount, in situ hybridization (wish) of 3-dpf zebrafish larvae (A) with cross (B) and longitudinal (C) sectioning of plastic embedded wish sample showing expression in epidermis, intestinal epithelium, and cells located in the brain. Bars: (full larvae) 300 µm; (sections) 100 µm. Immunostaining of ASC in 3-dpf Tg(krt4:GFP) larva (D). Optical cross section of lateral fin showing GFP expression in the EVL and ASC expression on both epidermal layers (D′). WT 3-dpf larva immunostained for ASC, together with nuclear envelope marker lamin and DAPI shows its nuclear and cytoplasmic localization (E). Immunostaining of 3-dpf Tg(mpeg1:EGFP) (F), Tg(lyz:DsRed2) (G), and Tg(spi1b:GAL4,UAS:TagRFP) (H) larvae showing expression of ASC in macrophages, neutrophils, and a single myeloid cell in the caudal hematopoietic tissue (H, white arrowhead). Bars: (full larvae) 300 µm; (all others) 30 µm.
ASC specks are highly intercrossed, filamentous structures whose clustering is altered by point mutations. (A–C) CLEM of high-pressure frozen 3 dpf Tg(HSE:asc-mKate2) larvae at 18 hphs. Low magnification electron micrograph (A, left) and overlay with red channel (A, right) imaged with light microscope. Black arrowhead shows location of speck. (B) Area of interest (red box) imaged with electron microscope. TEM tomography slice of the speck (B, black arrowhead) and overlay with 3D reconstruction of speck after manual tracking of individual filaments (B′). (C) Zoom in of three-dimensional reconstruction model. Entire TEM tomography stack and three-dimensional model are found in Video 4. Bars, 10 µm, unless otherwise indicated. (D) Results from phosphorylation-site analysis using the online tool GPS version 2.1.1, depicting Syk and JNK-specific, predicted phosphorylation sites in zebrafish ASC. Full results are found in Table S1. (E) Live imaging of larvae transiently expressing HSE:asc(4xmut)-mKate2, containing four missense mutations (T38A, Y152F, T160A, and T170A). (F) Single muscle cell in larvae transiently expressing either HSE:asc-mKate2, or HSE:asc(4xmut)-mKate2, or HSE:asc(Y152F)-mKate2. Bars, 30 µm.
ASC specks are highly intercrossed, filamentous structures whose clustering is altered by point mutations. (A–C) CLEM of high-pressure frozen 3 dpf Tg(HSE:asc-mKate2) larvae at 18 hphs. Low magnification electron micrograph (A, left) and overlay with red channel (A, right) imaged with light microscope. Black arrowhead shows location of speck. (B) Area of interest (red box) imaged with electron microscope. TEM tomography slice of the speck (B, black arrowhead) and overlay with 3D reconstruction of speck after manual tracking of individual filaments (B′). (C) Zoom in of three-dimensional reconstruction model. Entire TEM tomography stack and three-dimensional model are found in Video 4. Bars, 10 µm, unless otherwise indicated. (D) Results from phosphorylation-site analysis using the online tool GPS version 2.1.1, depicting Syk and JNK-specific, predicted phosphorylation sites in zebrafish ASC. Full results are found in Table S1. (E) Live imaging of larvae transiently expressing HSE:asc(4xmut)-mKate2, containing four missense mutations (T38A, Y152F, T160A, and T170A). (F) Single muscle cell in larvae transiently expressing either HSE:asc-mKate2, or HSE:asc(4xmut)-mKate2, or HSE:asc(Y152F)-mKate2. Bars, 30 µm.
