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machine learning software aivia  (Danaher Inc)


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    Structured Review

    Danaher Inc machine learning software aivia
    Figure 1. TIR-1 is expressed on lysosome-related organelles in C. elegans intestinal epithelial cells (A) Representative images of three fixed C. elegans TIR-1::3xFLAG animals at the L4 larval stage immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding differential interference contrast (DIC) images. Dotted boxes indicate higher magnifications. Dashed arrows in the middle panel indicate multiple vesicular structures. Dotted circles in the right panel highlight the lumen of vesicular structures. (B) Vector- and tir-1(RNAi)-treated TIR-1::3xFLAG animals were immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding DIC images. Dotted boxes indicate higher magnifications. (C) Representative images of three C. elegans TIR-1::3xFLAG animals co-immunostained with antibodies against FLAG (for TIR-1) or PGP-2 (for lysosome-related organelles). Images are three-dimensional reconstructions of z stacks. The last column shows TIR-1 (magenta) co-localized with PGP-2+ lysosome-related organelles (green). (D) Quantification of TIR-1 co-localization on PGP-2+ lysosome-related organelles (LROs) in animals stained only with anti-PGP-2 (control, n = 5 animals) or co- immunostained with both anti-FLAG and anti-PGP-2 by <t>Aivia</t> (n = 10 animals) (Leica). Box and whisker plots represent the median with minimum, second quartile, third quartile, and maximum indicated for each condition. *p < 0.05 (unpaired t test). (A)–(D) are representative of two independent experiments. Co-localization was quantified in (D) for animals that were intact after immunostaining. Scale bars as indicated. Source data for this figure is in Table S2. See also Figure S1.
    Machine Learning Software Aivia, supplied by Danaher Inc, used in various techniques. Bioz Stars score: 96/100, based on 186 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/aivia+machine+learning+software/Aivia+AI+Image+Analysis+Software/pm39299238-46-3-6
    Average 96 stars, based on 186 article reviews
    machine learning software aivia - by Bioz Stars, 2026-09
    96/100 stars

    Images

    1) Product Images from "Intestinal immunity in C. elegans is activated by pathogen effector-triggered aggregation of the guard protein TIR-1 on lysosome-related organelles."

    Article Title: Intestinal immunity in C. elegans is activated by pathogen effector-triggered aggregation of the guard protein TIR-1 on lysosome-related organelles.

    Journal: Immunity

    doi: 10.1016/j.immuni.2024.08.013

    Figure 1. TIR-1 is expressed on lysosome-related organelles in C. elegans intestinal epithelial cells (A) Representative images of three fixed C. elegans TIR-1::3xFLAG animals at the L4 larval stage immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding differential interference contrast (DIC) images. Dotted boxes indicate higher magnifications. Dashed arrows in the middle panel indicate multiple vesicular structures. Dotted circles in the right panel highlight the lumen of vesicular structures. (B) Vector- and tir-1(RNAi)-treated TIR-1::3xFLAG animals were immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding DIC images. Dotted boxes indicate higher magnifications. (C) Representative images of three C. elegans TIR-1::3xFLAG animals co-immunostained with antibodies against FLAG (for TIR-1) or PGP-2 (for lysosome-related organelles). Images are three-dimensional reconstructions of z stacks. The last column shows TIR-1 (magenta) co-localized with PGP-2+ lysosome-related organelles (green). (D) Quantification of TIR-1 co-localization on PGP-2+ lysosome-related organelles (LROs) in animals stained only with anti-PGP-2 (control, n = 5 animals) or co- immunostained with both anti-FLAG and anti-PGP-2 by Aivia (n = 10 animals) (Leica). Box and whisker plots represent the median with minimum, second quartile, third quartile, and maximum indicated for each condition. *p < 0.05 (unpaired t test). (A)–(D) are representative of two independent experiments. Co-localization was quantified in (D) for animals that were intact after immunostaining. Scale bars as indicated. Source data for this figure is in Table S2. See also Figure S1.
    Figure Legend Snippet: Figure 1. TIR-1 is expressed on lysosome-related organelles in C. elegans intestinal epithelial cells (A) Representative images of three fixed C. elegans TIR-1::3xFLAG animals at the L4 larval stage immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding differential interference contrast (DIC) images. Dotted boxes indicate higher magnifications. Dashed arrows in the middle panel indicate multiple vesicular structures. Dotted circles in the right panel highlight the lumen of vesicular structures. (B) Vector- and tir-1(RNAi)-treated TIR-1::3xFLAG animals were immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding DIC images. Dotted boxes indicate higher magnifications. (C) Representative images of three C. elegans TIR-1::3xFLAG animals co-immunostained with antibodies against FLAG (for TIR-1) or PGP-2 (for lysosome-related organelles). Images are three-dimensional reconstructions of z stacks. The last column shows TIR-1 (magenta) co-localized with PGP-2+ lysosome-related organelles (green). (D) Quantification of TIR-1 co-localization on PGP-2+ lysosome-related organelles (LROs) in animals stained only with anti-PGP-2 (control, n = 5 animals) or co- immunostained with both anti-FLAG and anti-PGP-2 by Aivia (n = 10 animals) (Leica). Box and whisker plots represent the median with minimum, second quartile, third quartile, and maximum indicated for each condition. *p < 0.05 (unpaired t test). (A)–(D) are representative of two independent experiments. Co-localization was quantified in (D) for animals that were intact after immunostaining. Scale bars as indicated. Source data for this figure is in Table S2. See also Figure S1.

    Techniques Used: Plasmid Preparation, Staining, Control, Whisker Assay, Immunostaining

    Figure 6. Pyocyanin triggers aggregation of TIR-1 on lysosome-related organelles and activates p38 PMK-1 intestinal immunity (A) Representative immunoblot of whole-cell lysates isolated from wild-type and TIR-1DHD mutants treated with the indicated phenazines for 4–6 h and probed with anti-phospho PMK-1, anti-total PMK-1, and anti-⍺-tubulin antibodies. (B) Densitometric quantification of conditions in (A) (n = 2). Data are mean of replicates with error bars giving SEM. *p < 0.05 (two-way ANOVA with Tukey’s multiple comparisons test). (C) Representative immunoblot of whole-cell lysates isolated from wild-type and pgp-2(kx48) mutants treated with or without PYO (200 mM) for 2–4 h and probed with anti-phospho PMK-1, anti-total PMK-1, and anti-⍺-tubulin antibodies. (D) Densitometric quantification of conditions in (C) (n = 3). Data are mean of replicates with error bars giving SEM. *p < 0.05 (two-way ANOVA with Sı´da´ k’s multiple comparisons test). (E) Representative pyocyanin toxicity fast-kill assay of wild-type, TIR-1DHD, and pgp-2(kx48) mutants. The difference between the wild type and the other ge- notypes is significant (*p < 0.05, log-rank test) (n = 3). (F) Mean lifespans with SEM for (E). *p < 0.05, log-rank test. (G) Representative images of animals treated with solvent control or PYO (200 mM) co-immunostained with anti-FLAG (for TIR-1) and anti-PGP-2. The last column represents the Aivia (Leica) render of TIR-1 and PGP-2 co-localization. (H) Quantification of TIR-1 puncta size on PGP-2+ vesicles in co-immunostained animals in the presence or absence of PYO. Each data point represents the average TIR-1 puncta size co-localized with a single PGP-2+ vesicle. Five vesicles were randomly identified per animal in five animals across two independent trials (n = 25). *p < 0.05 (unpaired t test). All phenazines were used at 200 mM except for 1-HP (20 mM). Scale bars as indicated. Mean lifespans and statistics for all replicates are in Table S1. Source data for this figure are in Table S2. See also Figure S6.
    Figure Legend Snippet: Figure 6. Pyocyanin triggers aggregation of TIR-1 on lysosome-related organelles and activates p38 PMK-1 intestinal immunity (A) Representative immunoblot of whole-cell lysates isolated from wild-type and TIR-1DHD mutants treated with the indicated phenazines for 4–6 h and probed with anti-phospho PMK-1, anti-total PMK-1, and anti-⍺-tubulin antibodies. (B) Densitometric quantification of conditions in (A) (n = 2). Data are mean of replicates with error bars giving SEM. *p < 0.05 (two-way ANOVA with Tukey’s multiple comparisons test). (C) Representative immunoblot of whole-cell lysates isolated from wild-type and pgp-2(kx48) mutants treated with or without PYO (200 mM) for 2–4 h and probed with anti-phospho PMK-1, anti-total PMK-1, and anti-⍺-tubulin antibodies. (D) Densitometric quantification of conditions in (C) (n = 3). Data are mean of replicates with error bars giving SEM. *p < 0.05 (two-way ANOVA with Sı´da´ k’s multiple comparisons test). (E) Representative pyocyanin toxicity fast-kill assay of wild-type, TIR-1DHD, and pgp-2(kx48) mutants. The difference between the wild type and the other ge- notypes is significant (*p < 0.05, log-rank test) (n = 3). (F) Mean lifespans with SEM for (E). *p < 0.05, log-rank test. (G) Representative images of animals treated with solvent control or PYO (200 mM) co-immunostained with anti-FLAG (for TIR-1) and anti-PGP-2. The last column represents the Aivia (Leica) render of TIR-1 and PGP-2 co-localization. (H) Quantification of TIR-1 puncta size on PGP-2+ vesicles in co-immunostained animals in the presence or absence of PYO. Each data point represents the average TIR-1 puncta size co-localized with a single PGP-2+ vesicle. Five vesicles were randomly identified per animal in five animals across two independent trials (n = 25). *p < 0.05 (unpaired t test). All phenazines were used at 200 mM except for 1-HP (20 mM). Scale bars as indicated. Mean lifespans and statistics for all replicates are in Table S1. Source data for this figure are in Table S2. See also Figure S6.

    Techniques Used: Western Blot, Isolation, Solvent, Control

    Related Articles

    Staining:

    Article Title: Interleukin-6 as a keystone cytokine in experimental rat models of chemotherapy-induced peripheral neurotoxicity.
    Article Snippet: .. CD68 immunoreactive area (brown staining) was identified and measured using the “Pixel Classifier” function of the Aivia image analysis software (Leica Microsystems, Bellevue, WA). ..

    Article Title: MI-181 enhances ciliation and cilia length in a cigarette smoke exposed airway epithelial model.
    Article Snippet: .. To do this, Zstacks of 100μm x 100μm images of ALI cultures from each donor and condition that had been stained for acetylated tubulin were analyzed using Leica Aivia artificial intelligence (AI) image analysis software to generate ACCEPTED MANUSCRIPT AR TIC LE IN PR ES S ARTICLE IN PRESS 3D images of the cilia on multiciliated cells (Fig. 3a-c). ..

    Article Title: MI-181 enhances ciliation and cilia length in a cigarette smoke exposed airway epithelial model
    Article Snippet: .. To do this, Z-stacks of 100 μm x 100 μm images of ALI cultures from each donor and condition that had been stained for acetylated tubulin were analyzed using Leica Aivia artificial intelligence (AI) image analysis software to generate 3D images of the cilia on multiciliated cells (Fig. a-c). ..

    Article Title: Interleukin-6 as a keystone cytokine in experimental rat models of chemotherapy-induced peripheral neurotoxicity
    Article Snippet: .. CD68 immunoreactive area (brown staining) was identified and measured using the “Pixel Classifier” function of the Aivia image analysis software (Leica Microsystems, Bellevue, WA). ..

    Article Title: Nicorandil attenuates thioacetamide induced liver fibrosis via AMPK, SIRT1 and HIF1α mediated cellular energy homeostasis
    Article Snippet: Microscopic evaluation was performed using an Olympus BX50 light microscope (Olympus, Japan), and digital images were captured with an Olympus imaging system at a magnification of 400×. .. Masson Trichrome stained hepatic sections were analyzed using image analysis software (LEICA microsystems (LAS version 3.8.0 (build:878), Leica Ltd) image analyzer computer system) at Cytology and Histology Department, Faculty of Veterinary Medicine, Cairo University. ..

    Software:

    Article Title: Interleukin-6 as a keystone cytokine in experimental rat models of chemotherapy-induced peripheral neurotoxicity.
    Article Snippet: .. CD68 immunoreactive area (brown staining) was identified and measured using the “Pixel Classifier” function of the Aivia image analysis software (Leica Microsystems, Bellevue, WA). ..

    Article Title: MI-181 enhances ciliation and cilia length in a cigarette smoke exposed airway epithelial model.
    Article Snippet: .. To do this, Zstacks of 100μm x 100μm images of ALI cultures from each donor and condition that had been stained for acetylated tubulin were analyzed using Leica Aivia artificial intelligence (AI) image analysis software to generate ACCEPTED MANUSCRIPT AR TIC LE IN PR ES S ARTICLE IN PRESS 3D images of the cilia on multiciliated cells (Fig. 3a-c). ..

    Article Title: Nicorandil attenuates thioacetamide induced liver fibrosis via AMPK, SIRT1 and HIF1α mediated cellular energy homeostasis
    Article Snippet: .. Anti-COX2 immunohistochemical sections were used to measure the mean area percent in different slides ( n = 5 fields/group) using image analysis software (LEICA microsystems (LAS version 3.8.0 (build:878), Leica Ltd) image analyzer computer system) at Cytology and Histology Department, Faculty of Veterinary Medicine, Cairo University. .. Data were statistically analyzed using SPSS software, version 22.0 (IBM Corp., Armonk, NY, USA).

    Article Title: MI-181 enhances ciliation and cilia length in a cigarette smoke exposed airway epithelial model
    Article Snippet: .. To do this, Z-stacks of 100 μm x 100 μm images of ALI cultures from each donor and condition that had been stained for acetylated tubulin were analyzed using Leica Aivia artificial intelligence (AI) image analysis software to generate 3D images of the cilia on multiciliated cells (Fig. a-c). ..

    Article Title: Interleukin-6 as a keystone cytokine in experimental rat models of chemotherapy-induced peripheral neurotoxicity
    Article Snippet: .. CD68 immunoreactive area (brown staining) was identified and measured using the “Pixel Classifier” function of the Aivia image analysis software (Leica Microsystems, Bellevue, WA). ..

    Article Title: Identification, Isolation, and In Vitro Culture Trials of Ovarian Germ Stem Cells from Different Teleost Fish Species
    Article Snippet: .. Images of H-E-stained slides were captured using a 20× objective with a digital camera (DFC 420; Leica Microsystems, Cambridge, UK) mounted on a light microscope and analyzed using an image-analysis software (Leica Application Suite X, version 5.1.0.25446, Wetzlar, Germany). ..

    Immunohistochemical staining:

    Article Title: Nicorandil attenuates thioacetamide induced liver fibrosis via AMPK, SIRT1 and HIF1α mediated cellular energy homeostasis
    Article Snippet: .. Anti-COX2 immunohistochemical sections were used to measure the mean area percent in different slides ( n = 5 fields/group) using image analysis software (LEICA microsystems (LAS version 3.8.0 (build:878), Leica Ltd) image analyzer computer system) at Cytology and Histology Department, Faculty of Veterinary Medicine, Cairo University. .. Data were statistically analyzed using SPSS software, version 22.0 (IBM Corp., Armonk, NY, USA).

    Light Microscopy:

    Article Title: Identification, Isolation, and In Vitro Culture Trials of Ovarian Germ Stem Cells from Different Teleost Fish Species
    Article Snippet: .. Images of H-E-stained slides were captured using a 20× objective with a digital camera (DFC 420; Leica Microsystems, Cambridge, UK) mounted on a light microscope and analyzed using an image-analysis software (Leica Application Suite X, version 5.1.0.25446, Wetzlar, Germany). ..

    Fluorescence:

    Article Title: In vitro culture of cumulus–oocyte complexes from early antral follicles in prepubertal and adult ewes
    Article Snippet: .. Image analysis software (Leica LAS AF Lite) was used to record the mean fluorescence intensity values by analyzing three different areas of each oocyte. ..



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    Figure 1. TIR-1 is expressed on lysosome-related organelles in C. elegans intestinal epithelial cells (A) Representative images of three fixed C. elegans TIR-1::3xFLAG animals at the L4 larval stage immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding differential interference contrast (DIC) images. Dotted boxes indicate higher magnifications. Dashed arrows in the middle panel indicate multiple vesicular structures. Dotted circles in the right panel highlight the lumen of vesicular structures. (B) Vector- and tir-1(RNAi)-treated TIR-1::3xFLAG animals were immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding DIC images. Dotted boxes indicate higher magnifications. (C) Representative images of three C. elegans TIR-1::3xFLAG animals co-immunostained with antibodies against FLAG (for TIR-1) or PGP-2 (for lysosome-related organelles). Images are three-dimensional reconstructions of z stacks. The last column shows TIR-1 (magenta) co-localized with PGP-2+ lysosome-related organelles (green). (D) Quantification of TIR-1 co-localization on PGP-2+ lysosome-related organelles (LROs) in animals stained only with anti-PGP-2 (control, n = 5 animals) or co- immunostained with both anti-FLAG and anti-PGP-2 by <t>Aivia</t> (n = 10 animals) (Leica). Box and whisker plots represent the median with minimum, second quartile, third quartile, and maximum indicated for each condition. *p < 0.05 (unpaired t test). (A)–(D) are representative of two independent experiments. Co-localization was quantified in (D) for animals that were intact after immunostaining. Scale bars as indicated. Source data for this figure is in Table S2. See also Figure S1.
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    Figure 1. TIR-1 is expressed on lysosome-related organelles in C. elegans intestinal epithelial cells (A) Representative images of three fixed C. elegans TIR-1::3xFLAG animals at the L4 larval stage immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding differential interference contrast (DIC) images. Dotted boxes indicate higher magnifications. Dashed arrows in the middle panel indicate multiple vesicular structures. Dotted circles in the right panel highlight the lumen of vesicular structures. (B) Vector- and tir-1(RNAi)-treated TIR-1::3xFLAG animals were immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding DIC images. Dotted boxes indicate higher magnifications. (C) Representative images of three C. elegans TIR-1::3xFLAG animals co-immunostained with antibodies against FLAG (for TIR-1) or PGP-2 (for lysosome-related organelles). Images are three-dimensional reconstructions of z stacks. The last column shows TIR-1 (magenta) co-localized with PGP-2+ lysosome-related organelles (green). (D) Quantification of TIR-1 co-localization on PGP-2+ lysosome-related organelles (LROs) in animals stained only with anti-PGP-2 (control, n = 5 animals) or co- immunostained with both anti-FLAG and anti-PGP-2 by <t>Aivia</t> (n = 10 animals) (Leica). Box and whisker plots represent the median with minimum, second quartile, third quartile, and maximum indicated for each condition. *p < 0.05 (unpaired t test). (A)–(D) are representative of two independent experiments. Co-localization was quantified in (D) for animals that were intact after immunostaining. Scale bars as indicated. Source data for this figure is in Table S2. See also Figure S1.
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    Figure 1. TIR-1 is expressed on lysosome-related organelles in C. elegans intestinal epithelial cells (A) Representative images of three fixed C. elegans TIR-1::3xFLAG animals at the L4 larval stage immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding differential interference contrast (DIC) images. Dotted boxes indicate higher magnifications. Dashed arrows in the middle panel indicate multiple vesicular structures. Dotted circles in the right panel highlight the lumen of vesicular structures. (B) Vector- and tir-1(RNAi)-treated TIR-1::3xFLAG animals were immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding DIC images. Dotted boxes indicate higher magnifications. (C) Representative images of three C. elegans TIR-1::3xFLAG animals co-immunostained with antibodies against FLAG (for TIR-1) or PGP-2 (for lysosome-related organelles). Images are three-dimensional reconstructions of z stacks. The last column shows TIR-1 (magenta) co-localized with PGP-2+ lysosome-related organelles (green). (D) Quantification of TIR-1 co-localization on PGP-2+ lysosome-related organelles (LROs) in animals stained only with anti-PGP-2 (control, n = 5 animals) or co- immunostained with both anti-FLAG and anti-PGP-2 by <t>Aivia</t> (n = 10 animals) (Leica). Box and whisker plots represent the median with minimum, second quartile, third quartile, and maximum indicated for each condition. *p < 0.05 (unpaired t test). (A)–(D) are representative of two independent experiments. Co-localization was quantified in (D) for animals that were intact after immunostaining. Scale bars as indicated. Source data for this figure is in Table S2. See also Figure S1.
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    Figure 1. TIR-1 is expressed on lysosome-related organelles in C. elegans intestinal epithelial cells (A) Representative images of three fixed C. elegans TIR-1::3xFLAG animals at the L4 larval stage immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding differential interference contrast (DIC) images. Dotted boxes indicate higher magnifications. Dashed arrows in the middle panel indicate multiple vesicular structures. Dotted circles in the right panel highlight the lumen of vesicular structures. (B) Vector- and tir-1(RNAi)-treated TIR-1::3xFLAG animals were immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding DIC images. Dotted boxes indicate higher magnifications. (C) Representative images of three C. elegans TIR-1::3xFLAG animals co-immunostained with antibodies against FLAG (for TIR-1) or PGP-2 (for lysosome-related organelles). Images are three-dimensional reconstructions of z stacks. The last column shows TIR-1 (magenta) co-localized with PGP-2+ lysosome-related organelles (green). (D) Quantification of TIR-1 co-localization on PGP-2+ lysosome-related organelles (LROs) in animals stained only with anti-PGP-2 (control, n = 5 animals) or co- immunostained with both anti-FLAG and anti-PGP-2 by <t>Aivia</t> (n = 10 animals) (Leica). Box and whisker plots represent the median with minimum, second quartile, third quartile, and maximum indicated for each condition. *p < 0.05 (unpaired t test). (A)–(D) are representative of two independent experiments. Co-localization was quantified in (D) for animals that were intact after immunostaining. Scale bars as indicated. Source data for this figure is in Table S2. See also Figure S1.
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    Image Search Results


    Figure 1. TIR-1 is expressed on lysosome-related organelles in C. elegans intestinal epithelial cells (A) Representative images of three fixed C. elegans TIR-1::3xFLAG animals at the L4 larval stage immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding differential interference contrast (DIC) images. Dotted boxes indicate higher magnifications. Dashed arrows in the middle panel indicate multiple vesicular structures. Dotted circles in the right panel highlight the lumen of vesicular structures. (B) Vector- and tir-1(RNAi)-treated TIR-1::3xFLAG animals were immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding DIC images. Dotted boxes indicate higher magnifications. (C) Representative images of three C. elegans TIR-1::3xFLAG animals co-immunostained with antibodies against FLAG (for TIR-1) or PGP-2 (for lysosome-related organelles). Images are three-dimensional reconstructions of z stacks. The last column shows TIR-1 (magenta) co-localized with PGP-2+ lysosome-related organelles (green). (D) Quantification of TIR-1 co-localization on PGP-2+ lysosome-related organelles (LROs) in animals stained only with anti-PGP-2 (control, n = 5 animals) or co- immunostained with both anti-FLAG and anti-PGP-2 by Aivia (n = 10 animals) (Leica). Box and whisker plots represent the median with minimum, second quartile, third quartile, and maximum indicated for each condition. *p < 0.05 (unpaired t test). (A)–(D) are representative of two independent experiments. Co-localization was quantified in (D) for animals that were intact after immunostaining. Scale bars as indicated. Source data for this figure is in Table S2. See also Figure S1.

    Journal: Immunity

    Article Title: Intestinal immunity in C. elegans is activated by pathogen effector-triggered aggregation of the guard protein TIR-1 on lysosome-related organelles.

    doi: 10.1016/j.immuni.2024.08.013

    Figure Lengend Snippet: Figure 1. TIR-1 is expressed on lysosome-related organelles in C. elegans intestinal epithelial cells (A) Representative images of three fixed C. elegans TIR-1::3xFLAG animals at the L4 larval stage immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding differential interference contrast (DIC) images. Dotted boxes indicate higher magnifications. Dashed arrows in the middle panel indicate multiple vesicular structures. Dotted circles in the right panel highlight the lumen of vesicular structures. (B) Vector- and tir-1(RNAi)-treated TIR-1::3xFLAG animals were immunostained with an anti-FLAG antibody (for TIR-1) and DAPI. Insets in the left panel represent corresponding DIC images. Dotted boxes indicate higher magnifications. (C) Representative images of three C. elegans TIR-1::3xFLAG animals co-immunostained with antibodies against FLAG (for TIR-1) or PGP-2 (for lysosome-related organelles). Images are three-dimensional reconstructions of z stacks. The last column shows TIR-1 (magenta) co-localized with PGP-2+ lysosome-related organelles (green). (D) Quantification of TIR-1 co-localization on PGP-2+ lysosome-related organelles (LROs) in animals stained only with anti-PGP-2 (control, n = 5 animals) or co- immunostained with both anti-FLAG and anti-PGP-2 by Aivia (n = 10 animals) (Leica). Box and whisker plots represent the median with minimum, second quartile, third quartile, and maximum indicated for each condition. *p < 0.05 (unpaired t test). (A)–(D) are representative of two independent experiments. Co-localization was quantified in (D) for animals that were intact after immunostaining. Scale bars as indicated. Source data for this figure is in Table S2. See also Figure S1.

    Article Snippet: We used the machine-learning software Aivia (Leica) to quantify the co-localization between TIR-1+ and PGP-2+ vesicles in three dimensions (see STAR Methods).

    Techniques: Plasmid Preparation, Staining, Control, Whisker Assay, Immunostaining

    Figure 6. Pyocyanin triggers aggregation of TIR-1 on lysosome-related organelles and activates p38 PMK-1 intestinal immunity (A) Representative immunoblot of whole-cell lysates isolated from wild-type and TIR-1DHD mutants treated with the indicated phenazines for 4–6 h and probed with anti-phospho PMK-1, anti-total PMK-1, and anti-⍺-tubulin antibodies. (B) Densitometric quantification of conditions in (A) (n = 2). Data are mean of replicates with error bars giving SEM. *p < 0.05 (two-way ANOVA with Tukey’s multiple comparisons test). (C) Representative immunoblot of whole-cell lysates isolated from wild-type and pgp-2(kx48) mutants treated with or without PYO (200 mM) for 2–4 h and probed with anti-phospho PMK-1, anti-total PMK-1, and anti-⍺-tubulin antibodies. (D) Densitometric quantification of conditions in (C) (n = 3). Data are mean of replicates with error bars giving SEM. *p < 0.05 (two-way ANOVA with Sı´da´ k’s multiple comparisons test). (E) Representative pyocyanin toxicity fast-kill assay of wild-type, TIR-1DHD, and pgp-2(kx48) mutants. The difference between the wild type and the other ge- notypes is significant (*p < 0.05, log-rank test) (n = 3). (F) Mean lifespans with SEM for (E). *p < 0.05, log-rank test. (G) Representative images of animals treated with solvent control or PYO (200 mM) co-immunostained with anti-FLAG (for TIR-1) and anti-PGP-2. The last column represents the Aivia (Leica) render of TIR-1 and PGP-2 co-localization. (H) Quantification of TIR-1 puncta size on PGP-2+ vesicles in co-immunostained animals in the presence or absence of PYO. Each data point represents the average TIR-1 puncta size co-localized with a single PGP-2+ vesicle. Five vesicles were randomly identified per animal in five animals across two independent trials (n = 25). *p < 0.05 (unpaired t test). All phenazines were used at 200 mM except for 1-HP (20 mM). Scale bars as indicated. Mean lifespans and statistics for all replicates are in Table S1. Source data for this figure are in Table S2. See also Figure S6.

    Journal: Immunity

    Article Title: Intestinal immunity in C. elegans is activated by pathogen effector-triggered aggregation of the guard protein TIR-1 on lysosome-related organelles.

    doi: 10.1016/j.immuni.2024.08.013

    Figure Lengend Snippet: Figure 6. Pyocyanin triggers aggregation of TIR-1 on lysosome-related organelles and activates p38 PMK-1 intestinal immunity (A) Representative immunoblot of whole-cell lysates isolated from wild-type and TIR-1DHD mutants treated with the indicated phenazines for 4–6 h and probed with anti-phospho PMK-1, anti-total PMK-1, and anti-⍺-tubulin antibodies. (B) Densitometric quantification of conditions in (A) (n = 2). Data are mean of replicates with error bars giving SEM. *p < 0.05 (two-way ANOVA with Tukey’s multiple comparisons test). (C) Representative immunoblot of whole-cell lysates isolated from wild-type and pgp-2(kx48) mutants treated with or without PYO (200 mM) for 2–4 h and probed with anti-phospho PMK-1, anti-total PMK-1, and anti-⍺-tubulin antibodies. (D) Densitometric quantification of conditions in (C) (n = 3). Data are mean of replicates with error bars giving SEM. *p < 0.05 (two-way ANOVA with Sı´da´ k’s multiple comparisons test). (E) Representative pyocyanin toxicity fast-kill assay of wild-type, TIR-1DHD, and pgp-2(kx48) mutants. The difference between the wild type and the other ge- notypes is significant (*p < 0.05, log-rank test) (n = 3). (F) Mean lifespans with SEM for (E). *p < 0.05, log-rank test. (G) Representative images of animals treated with solvent control or PYO (200 mM) co-immunostained with anti-FLAG (for TIR-1) and anti-PGP-2. The last column represents the Aivia (Leica) render of TIR-1 and PGP-2 co-localization. (H) Quantification of TIR-1 puncta size on PGP-2+ vesicles in co-immunostained animals in the presence or absence of PYO. Each data point represents the average TIR-1 puncta size co-localized with a single PGP-2+ vesicle. Five vesicles were randomly identified per animal in five animals across two independent trials (n = 25). *p < 0.05 (unpaired t test). All phenazines were used at 200 mM except for 1-HP (20 mM). Scale bars as indicated. Mean lifespans and statistics for all replicates are in Table S1. Source data for this figure are in Table S2. See also Figure S6.

    Article Snippet: We used the machine-learning software Aivia (Leica) to quantify the co-localization between TIR-1+ and PGP-2+ vesicles in three dimensions (see STAR Methods).

    Techniques: Western Blot, Isolation, Solvent, Control