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jnj 55308942  (MedChemExpress)
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Meningeal stromal cell‐derived IL‐33 and ATP coordinate antiviral defense. A) Maximal intensity projection of dural meninges stained for stromal cells (green), IL‐33 (red), LCMV (white) and nuclei (blue) at 6 dpi. Scale bar, 2000 µm (left, whole mount); 100 µm (right). n = 6 (mice). B) The density of meningeal IL‐33 + stromal cells (A). n = 20 (clusters, from 6 mice). C–H) Primary meningeal stromal cells were cultured. C) Staining of IL‐33 (red), nuclei (blue), and LCMV (white) post LCMV infection. MOI = 2:1. Scale bar, 200 µm. n = 6–7 (views). D) The percentage of IL‐33 high cells (C). E) The percentage of IL‐33 high cells treated with medium alone (NC), LCMV, LCMV + 2‐APB at 1 dpi. MOI = 1:1. n = 25‐28 (views). F) The percentage of IL‐33 high cells treated with medium alone (NC), or ionomycin + PMA for 1 day. n = 24‐36 (views). G) Heatmap of differentially expressed genes at 1 dpi. MOI = 2:1. n = 3 (wells, each well from 2 mice). H) The extracellular ATP level after LCMV infection. MOI = 2:1. n = 2‐3 (wells, each well from 2 mice). I) Live cell imaging of calcium flux in PCMCs from Tpsb2‐CreERT2; Ai162 fl/+ mice stimulated with conditioned medium from virus‐infected stromal cells. Scale bar, 50 µm. n = 75–275 (cells). J) Quantification of (I). K) Live cell imaging of calcium flux in BMMCs from Tpsb2‐CreERT2; Ai162 fl/+ mice stimulated with IL‐33, ATP, or ionomycin + PMA. Scale bar, 50 µm. n = 156‐356 (cells). L) Quantification of (K). M) Live cell imaging of calcium flux in the meninges from Tpsb2‐CreERT2; Ai162 fl/+ mice with the stimulation of ATP ex vivo . Scale bar, 200 µm. n = 33 (cells). N) Quantification of (M). O) Meningeal immune cell subset counts treated with ATP receptor antagonist (AZD9056, JNJ55308942, suramin sodium salt) at 6 dpi. n = 7–8 (mice). Data are presented as mean ± SEM and p values were calculated by two‐tailed, paired Student's t ‐test (B), two‐tailed, unpaired Student's t ‐test (D–F, H, O), Wald test from DESeq2 (G).
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jnj-55308942  (Janssen)
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Meningeal stromal cell‐derived IL‐33 and ATP coordinate antiviral defense. A) Maximal intensity projection of dural meninges stained for stromal cells (green), IL‐33 (red), LCMV (white) and nuclei (blue) at 6 dpi. Scale bar, 2000 µm (left, whole mount); 100 µm (right). n = 6 (mice). B) The density of meningeal IL‐33 + stromal cells (A). n = 20 (clusters, from 6 mice). C–H) Primary meningeal stromal cells were cultured. C) Staining of IL‐33 (red), nuclei (blue), and LCMV (white) post LCMV infection. MOI = 2:1. Scale bar, 200 µm. n = 6–7 (views). D) The percentage of IL‐33 high cells (C). E) The percentage of IL‐33 high cells treated with medium alone (NC), LCMV, LCMV + 2‐APB at 1 dpi. MOI = 1:1. n = 25‐28 (views). F) The percentage of IL‐33 high cells treated with medium alone (NC), or ionomycin + PMA for 1 day. n = 24‐36 (views). G) Heatmap of differentially expressed genes at 1 dpi. MOI = 2:1. n = 3 (wells, each well from 2 mice). H) The extracellular ATP level after LCMV infection. MOI = 2:1. n = 2‐3 (wells, each well from 2 mice). I) Live cell imaging of calcium flux in PCMCs from Tpsb2‐CreERT2; Ai162 fl/+ mice stimulated with conditioned medium from virus‐infected stromal cells. Scale bar, 50 µm. n = 75–275 (cells). J) Quantification of (I). K) Live cell imaging of calcium flux in BMMCs from Tpsb2‐CreERT2; Ai162 fl/+ mice stimulated with IL‐33, ATP, or ionomycin + PMA. Scale bar, 50 µm. n = 156‐356 (cells). L) Quantification of (K). M) Live cell imaging of calcium flux in the meninges from Tpsb2‐CreERT2; Ai162 fl/+ mice with the stimulation of ATP ex vivo . Scale bar, 200 µm. n = 33 (cells). N) Quantification of (M). O) Meningeal immune cell subset counts treated with ATP receptor antagonist (AZD9056, JNJ55308942, suramin sodium salt) at 6 dpi. n = 7–8 (mice). Data are presented as mean ± SEM and p values were calculated by two‐tailed, paired Student's t ‐test (B), two‐tailed, unpaired Student's t ‐test (D–F, H, O), Wald test from DESeq2 (G).
Jnj 55308942, supplied by Janssen, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Meningeal stromal cell‐derived IL‐33 and ATP coordinate antiviral defense. A) Maximal intensity projection of dural meninges stained for stromal cells (green), IL‐33 (red), LCMV (white) and nuclei (blue) at 6 dpi. Scale bar, 2000 µm (left, whole mount); 100 µm (right). n = 6 (mice). B) The density of meningeal IL‐33 + stromal cells (A). n = 20 (clusters, from 6 mice). C–H) Primary meningeal stromal cells were cultured. C) Staining of IL‐33 (red), nuclei (blue), and LCMV (white) post LCMV infection. MOI = 2:1. Scale bar, 200 µm. n = 6–7 (views). D) The percentage of IL‐33 high cells (C). E) The percentage of IL‐33 high cells treated with medium alone (NC), LCMV, LCMV + 2‐APB at 1 dpi. MOI = 1:1. n = 25‐28 (views). F) The percentage of IL‐33 high cells treated with medium alone (NC), or ionomycin + PMA for 1 day. n = 24‐36 (views). G) Heatmap of differentially expressed genes at 1 dpi. MOI = 2:1. n = 3 (wells, each well from 2 mice). H) The extracellular ATP level after LCMV infection. MOI = 2:1. n = 2‐3 (wells, each well from 2 mice). I) Live cell imaging of calcium flux in PCMCs from Tpsb2‐CreERT2; Ai162 fl/+ mice stimulated with conditioned medium from virus‐infected stromal cells. Scale bar, 50 µm. n = 75–275 (cells). J) Quantification of (I). K) Live cell imaging of calcium flux in BMMCs from Tpsb2‐CreERT2; Ai162 fl/+ mice stimulated with IL‐33, ATP, or ionomycin + PMA. Scale bar, 50 µm. n = 156‐356 (cells). L) Quantification of (K). M) Live cell imaging of calcium flux in the meninges from Tpsb2‐CreERT2; Ai162 fl/+ mice with the stimulation of ATP ex vivo . Scale bar, 200 µm. n = 33 (cells). N) Quantification of (M). O) Meningeal immune cell subset counts treated with ATP receptor antagonist (AZD9056, JNJ55308942, suramin sodium salt) at 6 dpi. n = 7–8 (mice). Data are presented as mean ± SEM and p values were calculated by two‐tailed, paired Student's t ‐test (B), two‐tailed, unpaired Student's t ‐test (D–F, H, O), Wald test from DESeq2 (G).

Journal: Advanced Science

Article Title: Stromal Cell‐Mast Cell Communication Orchestrates Anti‐Viral Immunity in the Meninges

doi: 10.1002/advs.202514842

Figure Lengend Snippet: Meningeal stromal cell‐derived IL‐33 and ATP coordinate antiviral defense. A) Maximal intensity projection of dural meninges stained for stromal cells (green), IL‐33 (red), LCMV (white) and nuclei (blue) at 6 dpi. Scale bar, 2000 µm (left, whole mount); 100 µm (right). n = 6 (mice). B) The density of meningeal IL‐33 + stromal cells (A). n = 20 (clusters, from 6 mice). C–H) Primary meningeal stromal cells were cultured. C) Staining of IL‐33 (red), nuclei (blue), and LCMV (white) post LCMV infection. MOI = 2:1. Scale bar, 200 µm. n = 6–7 (views). D) The percentage of IL‐33 high cells (C). E) The percentage of IL‐33 high cells treated with medium alone (NC), LCMV, LCMV + 2‐APB at 1 dpi. MOI = 1:1. n = 25‐28 (views). F) The percentage of IL‐33 high cells treated with medium alone (NC), or ionomycin + PMA for 1 day. n = 24‐36 (views). G) Heatmap of differentially expressed genes at 1 dpi. MOI = 2:1. n = 3 (wells, each well from 2 mice). H) The extracellular ATP level after LCMV infection. MOI = 2:1. n = 2‐3 (wells, each well from 2 mice). I) Live cell imaging of calcium flux in PCMCs from Tpsb2‐CreERT2; Ai162 fl/+ mice stimulated with conditioned medium from virus‐infected stromal cells. Scale bar, 50 µm. n = 75–275 (cells). J) Quantification of (I). K) Live cell imaging of calcium flux in BMMCs from Tpsb2‐CreERT2; Ai162 fl/+ mice stimulated with IL‐33, ATP, or ionomycin + PMA. Scale bar, 50 µm. n = 156‐356 (cells). L) Quantification of (K). M) Live cell imaging of calcium flux in the meninges from Tpsb2‐CreERT2; Ai162 fl/+ mice with the stimulation of ATP ex vivo . Scale bar, 200 µm. n = 33 (cells). N) Quantification of (M). O) Meningeal immune cell subset counts treated with ATP receptor antagonist (AZD9056, JNJ55308942, suramin sodium salt) at 6 dpi. n = 7–8 (mice). Data are presented as mean ± SEM and p values were calculated by two‐tailed, paired Student's t ‐test (B), two‐tailed, unpaired Student's t ‐test (D–F, H, O), Wald test from DESeq2 (G).

Article Snippet: For ATP receptor blockade experiments, mice received intraperitoneal injections of 12.5 mg kg −1 AZD9056 (MCE, HY‐19427A), 12.5 mg kg −1 JNJ‐55308942 (MCE, HY‐123857), or 20 mg kg −1 suramin (MCE, HY‐B0879A) on days 1, 3, and 5 post‐infection.

Techniques: Derivative Assay, Staining, Cell Culture, Infection, Live Cell Imaging, Virus, Ex Vivo, Two Tailed Test