phenol red Search Results


hskmc growth medium  (Cell Applications Inc)
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Hskmc Growth Medium, supplied by Cell Applications Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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phenol red  (Beyotime)
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Beyotime phenol red
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cultrex pathclear bme gels  (R&D Systems)
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cultrex basement membrane extract  (R&D Systems)
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R&D Systems cultrex basement membrane extract
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rpmi 1640  (R&D Systems)
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R&D Systems rpmi 1640
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f 12 media  (R&D Systems)
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hyq pf  (Danaher Inc)
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matrigel domes  (R&D Systems)
94
R&D Systems matrigel domes
Human duodenal organoids were trypsinised into single cells and seeded onto culture plates pre-coated with <t>1:40</t> <t>Matrigel:PBS.</t> Cells were infected with S. flexneri ∆ ospc3 at an MOI of 100 for 4 h. CFUs in infected cells were enumerated ( a ), PI uptake was monitored using incucyte live cell imager and PI uptake was normalised per area confluency ( b ), images of monolayers taken 4 h post infection taken at 20x (i) scramble, uninfected; (ii) scramble, Shigella flexneri ∆ ospc3, (iii) GSDMD KD uninfected, (iv) GSDMD KD Shigella flexneri ∆ ospc3. Scale bars (indicting 200 micrometres) are depicted for each micrograph ( c ). Caspase-4 was measured in western blot of whole cell lysates ( d ), IL-18 secretion into the supernatant was measured by ELISA on cell free supernatants ( e ). A schematic of caspase-4 regulation ( f ). In ( f ) (i) LPS-induced caspase-4 activation (1) leads to activation of GSDMD and IL-18 (2). GSDMD pores lead to caspase-4 cleavage (3) and this stops IL-18 processing (4). In ( f )(ii), during conditions of reduced GSDMD –mediated plasma pore formation, caspase-4 activation (1) leads to IL-18 processing. A lack of efficient pore formation reduces inhibitory feedback on caspase-4 (3). This leads to continued activity of caspase-4 on IL-18 and IL-18 hypersecretion (4). All experiments are representative of at least 3 experiments. Data displayed as mean ± S.E.M. * p < 0.05, **** p < 0.001. Panel ( f ) created with Biorender.
Matrigel Domes, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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extracellular matrix ecm coating  (R&D Systems)
94
R&D Systems extracellular matrix ecm coating
Human duodenal organoids were trypsinised into single cells and seeded onto culture plates pre-coated with <t>1:40</t> <t>Matrigel:PBS.</t> Cells were infected with S. flexneri ∆ ospc3 at an MOI of 100 for 4 h. CFUs in infected cells were enumerated ( a ), PI uptake was monitored using incucyte live cell imager and PI uptake was normalised per area confluency ( b ), images of monolayers taken 4 h post infection taken at 20x (i) scramble, uninfected; (ii) scramble, Shigella flexneri ∆ ospc3, (iii) GSDMD KD uninfected, (iv) GSDMD KD Shigella flexneri ∆ ospc3. Scale bars (indicting 200 micrometres) are depicted for each micrograph ( c ). Caspase-4 was measured in western blot of whole cell lysates ( d ), IL-18 secretion into the supernatant was measured by ELISA on cell free supernatants ( e ). A schematic of caspase-4 regulation ( f ). In ( f ) (i) LPS-induced caspase-4 activation (1) leads to activation of GSDMD and IL-18 (2). GSDMD pores lead to caspase-4 cleavage (3) and this stops IL-18 processing (4). In ( f )(ii), during conditions of reduced GSDMD –mediated plasma pore formation, caspase-4 activation (1) leads to IL-18 processing. A lack of efficient pore formation reduces inhibitory feedback on caspase-4 (3). This leads to continued activity of caspase-4 on IL-18 and IL-18 hypersecretion (4). All experiments are representative of at least 3 experiments. Data displayed as mean ± S.E.M. * p < 0.05, **** p < 0.001. Panel ( f ) created with Biorender.
Extracellular Matrix Ecm Coating, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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phenol red  (Santa Cruz Biotechnology)
93
Santa Cruz Biotechnology phenol red
Human duodenal organoids were trypsinised into single cells and seeded onto culture plates pre-coated with <t>1:40</t> <t>Matrigel:PBS.</t> Cells were infected with S. flexneri ∆ ospc3 at an MOI of 100 for 4 h. CFUs in infected cells were enumerated ( a ), PI uptake was monitored using incucyte live cell imager and PI uptake was normalised per area confluency ( b ), images of monolayers taken 4 h post infection taken at 20x (i) scramble, uninfected; (ii) scramble, Shigella flexneri ∆ ospc3, (iii) GSDMD KD uninfected, (iv) GSDMD KD Shigella flexneri ∆ ospc3. Scale bars (indicting 200 micrometres) are depicted for each micrograph ( c ). Caspase-4 was measured in western blot of whole cell lysates ( d ), IL-18 secretion into the supernatant was measured by ELISA on cell free supernatants ( e ). A schematic of caspase-4 regulation ( f ). In ( f ) (i) LPS-induced caspase-4 activation (1) leads to activation of GSDMD and IL-18 (2). GSDMD pores lead to caspase-4 cleavage (3) and this stops IL-18 processing (4). In ( f )(ii), during conditions of reduced GSDMD –mediated plasma pore formation, caspase-4 activation (1) leads to IL-18 processing. A lack of efficient pore formation reduces inhibitory feedback on caspase-4 (3). This leads to continued activity of caspase-4 on IL-18 and IL-18 hypersecretion (4). All experiments are representative of at least 3 experiments. Data displayed as mean ± S.E.M. * p < 0.05, **** p < 0.001. Panel ( f ) created with Biorender.
Phenol Red, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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type 2 cultrex rgf basement membrane extract bme  (R&D Systems)
94
R&D Systems type 2 cultrex rgf basement membrane extract bme
Human duodenal organoids were trypsinised into single cells and seeded onto culture plates pre-coated with <t>1:40</t> <t>Matrigel:PBS.</t> Cells were infected with S. flexneri ∆ ospc3 at an MOI of 100 for 4 h. CFUs in infected cells were enumerated ( a ), PI uptake was monitored using incucyte live cell imager and PI uptake was normalised per area confluency ( b ), images of monolayers taken 4 h post infection taken at 20x (i) scramble, uninfected; (ii) scramble, Shigella flexneri ∆ ospc3, (iii) GSDMD KD uninfected, (iv) GSDMD KD Shigella flexneri ∆ ospc3. Scale bars (indicting 200 micrometres) are depicted for each micrograph ( c ). Caspase-4 was measured in western blot of whole cell lysates ( d ), IL-18 secretion into the supernatant was measured by ELISA on cell free supernatants ( e ). A schematic of caspase-4 regulation ( f ). In ( f ) (i) LPS-induced caspase-4 activation (1) leads to activation of GSDMD and IL-18 (2). GSDMD pores lead to caspase-4 cleavage (3) and this stops IL-18 processing (4). In ( f )(ii), during conditions of reduced GSDMD –mediated plasma pore formation, caspase-4 activation (1) leads to IL-18 processing. A lack of efficient pore formation reduces inhibitory feedback on caspase-4 (3). This leads to continued activity of caspase-4 on IL-18 and IL-18 hypersecretion (4). All experiments are representative of at least 3 experiments. Data displayed as mean ± S.E.M. * p < 0.05, **** p < 0.001. Panel ( f ) created with Biorender.
Type 2 Cultrex Rgf Basement Membrane Extract Bme, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rpmi cultrex basement membrane extract type 3  (R&D Systems)
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R&D Systems rpmi cultrex basement membrane extract type 3
Human duodenal organoids were trypsinised into single cells and seeded onto culture plates pre-coated with <t>1:40</t> <t>Matrigel:PBS.</t> Cells were infected with S. flexneri ∆ ospc3 at an MOI of 100 for 4 h. CFUs in infected cells were enumerated ( a ), PI uptake was monitored using incucyte live cell imager and PI uptake was normalised per area confluency ( b ), images of monolayers taken 4 h post infection taken at 20x (i) scramble, uninfected; (ii) scramble, Shigella flexneri ∆ ospc3, (iii) GSDMD KD uninfected, (iv) GSDMD KD Shigella flexneri ∆ ospc3. Scale bars (indicting 200 micrometres) are depicted for each micrograph ( c ). Caspase-4 was measured in western blot of whole cell lysates ( d ), IL-18 secretion into the supernatant was measured by ELISA on cell free supernatants ( e ). A schematic of caspase-4 regulation ( f ). In ( f ) (i) LPS-induced caspase-4 activation (1) leads to activation of GSDMD and IL-18 (2). GSDMD pores lead to caspase-4 cleavage (3) and this stops IL-18 processing (4). In ( f )(ii), during conditions of reduced GSDMD –mediated plasma pore formation, caspase-4 activation (1) leads to IL-18 processing. A lack of efficient pore formation reduces inhibitory feedback on caspase-4 (3). This leads to continued activity of caspase-4 on IL-18 and IL-18 hypersecretion (4). All experiments are representative of at least 3 experiments. Data displayed as mean ± S.E.M. * p < 0.05, **** p < 0.001. Panel ( f ) created with Biorender.
Rpmi Cultrex Basement Membrane Extract Type 3, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


Human duodenal organoids were trypsinised into single cells and seeded onto culture plates pre-coated with 1:40 Matrigel:PBS. Cells were infected with S. flexneri ∆ ospc3 at an MOI of 100 for 4 h. CFUs in infected cells were enumerated ( a ), PI uptake was monitored using incucyte live cell imager and PI uptake was normalised per area confluency ( b ), images of monolayers taken 4 h post infection taken at 20x (i) scramble, uninfected; (ii) scramble, Shigella flexneri ∆ ospc3, (iii) GSDMD KD uninfected, (iv) GSDMD KD Shigella flexneri ∆ ospc3. Scale bars (indicting 200 micrometres) are depicted for each micrograph ( c ). Caspase-4 was measured in western blot of whole cell lysates ( d ), IL-18 secretion into the supernatant was measured by ELISA on cell free supernatants ( e ). A schematic of caspase-4 regulation ( f ). In ( f ) (i) LPS-induced caspase-4 activation (1) leads to activation of GSDMD and IL-18 (2). GSDMD pores lead to caspase-4 cleavage (3) and this stops IL-18 processing (4). In ( f )(ii), during conditions of reduced GSDMD –mediated plasma pore formation, caspase-4 activation (1) leads to IL-18 processing. A lack of efficient pore formation reduces inhibitory feedback on caspase-4 (3). This leads to continued activity of caspase-4 on IL-18 and IL-18 hypersecretion (4). All experiments are representative of at least 3 experiments. Data displayed as mean ± S.E.M. * p < 0.05, **** p < 0.001. Panel ( f ) created with Biorender.

Journal: Communications Biology

Article Title: Gasdermin-D pores induce an inactivating caspase-4 cleavage that limits IL-18 production in the intestinal epithelium

doi: 10.1038/s42003-025-08183-9

Figure Lengend Snippet: Human duodenal organoids were trypsinised into single cells and seeded onto culture plates pre-coated with 1:40 Matrigel:PBS. Cells were infected with S. flexneri ∆ ospc3 at an MOI of 100 for 4 h. CFUs in infected cells were enumerated ( a ), PI uptake was monitored using incucyte live cell imager and PI uptake was normalised per area confluency ( b ), images of monolayers taken 4 h post infection taken at 20x (i) scramble, uninfected; (ii) scramble, Shigella flexneri ∆ ospc3, (iii) GSDMD KD uninfected, (iv) GSDMD KD Shigella flexneri ∆ ospc3. Scale bars (indicting 200 micrometres) are depicted for each micrograph ( c ). Caspase-4 was measured in western blot of whole cell lysates ( d ), IL-18 secretion into the supernatant was measured by ELISA on cell free supernatants ( e ). A schematic of caspase-4 regulation ( f ). In ( f ) (i) LPS-induced caspase-4 activation (1) leads to activation of GSDMD and IL-18 (2). GSDMD pores lead to caspase-4 cleavage (3) and this stops IL-18 processing (4). In ( f )(ii), during conditions of reduced GSDMD –mediated plasma pore formation, caspase-4 activation (1) leads to IL-18 processing. A lack of efficient pore formation reduces inhibitory feedback on caspase-4 (3). This leads to continued activity of caspase-4 on IL-18 and IL-18 hypersecretion (4). All experiments are representative of at least 3 experiments. Data displayed as mean ± S.E.M. * p < 0.05, **** p < 0.001. Panel ( f ) created with Biorender.

Article Snippet: Tissue was minced until able to be passed through a 1 mL pipette tip then incubated in 2 mg/mL collagenase at 37 o C for 40 min. Crypt units were dissociated by vigorous pipetting then washed 3x in PBS containing 10% FBS and plated into 40 μL Matrigel domes (Cultrex PathClear reduced growth factor BME, type 2; R&D Systems) in a 24 well plate.

Techniques: Infection, Western Blot, Enzyme-linked Immunosorbent Assay, Activation Assay, Clinical Proteomics, Activity Assay