darapladib (TargetMol)
Structured Review

Darapladib, supplied by TargetMol, used in various techniques. Bioz Stars score: 94/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 94 stars, based on 2 article reviews
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1) Product Images from "Inhibiting ceramide synthase 5 expression in microglia decreases neuroinflammation after spinal cord injury"
Article Title: Inhibiting ceramide synthase 5 expression in microglia decreases neuroinflammation after spinal cord injury
Journal: Neural Regeneration Research
doi: 10.4103/NRR.NRR-D-23-01933
Figure Legend Snippet: Single-cell analysis of microglia in the acute/subacute phase after SCI. (A) UMAP plot of all 8251 microglia collected from uninjured and injured spinal cords at 3, 7, and 14 dpi. Cells are colored and annotated by cell subsets based on a combination of DEGs, canonical marker genes, and previously published spinal cord scRNA-seq datasets. (B) Heatmap showing the marker genes for different microglial subsets. (C) The proportion of different microglia subsets at four time points: uninjured and 3, 7, and 14 dpi. (D) Gene Ontology (GO) analysis of microglia in the MG1 subset. (E) GSVA analysis of microglial subsets. (F) The expression of sphingomyelin metabolism-related genes ( Cers5 , Pla2g7 ), pyroptosis-related genes ( Gsdmd , Nlrp3 , Casp1 ), and inflammation-related genes ( Nfkb1 , Il1b ) in different microglia subsets. The scRNA-seq dataset used in this figure was obtained from the Figshare database ( https://doi.org/10.6084/m9.figshare.17702045.v2 ). Casp1: Caspase 1; Cers5: ceramide synthase 5; DEG: differentially expressed gene; dpi: days post injury; GO: gene ontology; Gsdmd: gasdermin D; GSVA: gene set variation analysis; Il1b: interleukin 1β; Nfkb1: nuclear factor kappa-B 1; Nlrp3: NOD-like receptor thermal protein domain associated protein 3; Pla2g7: phospholipase a2 group VII; scRNA-seq: ScRNA-seq; UMAP: uniform manifold approximation and projection.
Techniques Used: Single-cell Analysis, Marker, Expressing
Figure Legend Snippet: CerS5 knockdown inhibits Pla2g7/p50 expression in the spinal cord after SCI. (A) Western blot of Pla2g7 and p50 expression levels in the SCI and Sham groups after treatment with siCerS5 ( n = 3). (B, C) Representative examples of immunofluorescence staining images used to analyze the percentage of Pla2g7 + or p50 + microglia out of all microglia in the SCI and Sham groups after treatment with si.CerS5 ( n = 5). There were significantly fewer Pla2g7 + Iba1 + cells and p50 + Iba1 + cells in the SCI + si.CerS5 group compared with the SCI and SCI + Si.NC groups. Iba1, green, CoraLite 488; Pla2g7/p50, red, CoraLite 594; DAPI, blue. Data are expressed as the mean ± SEM. ** P < 0.01 (one-way analysis of variance followed by Tukey’s multiple comparison test). CerS: Ceramide synthase; DAPI: 4′,6-diamidino-2-phenylindole; Iba1: ionized calcium binding adaptor molecule 1; p50: nuclear factor kappa-B p105/p50; SCI: spinal cord injury; si.CerS5: CerS5 siRNA.
Techniques Used: Knockdown, Expressing, Western Blot, Immunofluorescence, Staining, Comparison, Binding Assay
Figure Legend Snippet: Pla2g7 is essential for mediating the regulatory effects of CerS5 on pyroptosis. (A) Ceramide concentrations in the spinal cord in the SCI and Sham groups were detected by ELISA, and the data were normalized to the Shame group ( n = 3). (B) Western blot analysis of Pla2g7, p50, GSDMD, and GSDMD-N expression levels in BV2 cells after treatment with Darapladib (Pla2g7 inhibitor) ( n = 3). ** P < 0.01, vs . C16 ceramide. (C) Representative examples of immunofluorescence staining images used to quantify GSDMD + Iba1 + cells after treatment with Darapladib ( n = 5). There were significantly fewer GSDMD + Iba1 + cells in the C16 ceramide + Darapladib group than in the C16 ceramide group. Iba1, green, CoraLite 488; GSDMD, red, CoraLite 594; DAPI, blue. Scale bar: 100 μm. (D) Western blot analysis of the expression levels of proteins associated with the classical NLRP3 pyroptosis pathway in BV2 cells after treatment with siCerS5 or Darapladib ( n = 3). Data are expressed as the mean ± SEM. * P < 0.05, ** P < 0.01 (one-way analysis of variance followed by Tukey’s multiple comparison test). ASC: Apoptosis associated speck like protein containing CARD; CerS: ceramide synthase; DAPI: 4′,6-diamidino-2-phenylindole; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GSDMD: gasdermin D; GSDMD-N: N-terminal GSDMD; Iba1: ionized calcium binding adaptor molecule 1; NLRP3: NOD-like receptor thermal protein domain associated protein 3; SCI: spinal cord injury; si.CerS5: CerS5 siRNA.
Techniques Used: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Immunofluorescence, Staining, Comparison, Binding Assay
Figure Legend Snippet: Darapladib inhibits microglial pyroptosis in the spinal cord after SCI. (A) Representative examples of immunofluorescence staining images used to quantify GSDMD + Iba1 + cells in the SCI and Sham groups after treatment with Darapladib ( n = 5). There were significantly fewer GSDMD + Iba1 + cells in the SCI + Darapladib group compared with the SCI and SCI + Vehicle groups. Iba1, green, CoraLite 488; GSDMD, red, CoraLite 594; DAPI, blue. Scale bars: 100 μm (upper); 20 μm (lower). (B) Western blot analysis of the expression levels of Pla2g7 and proteins associated with the classical NLRP3 pyroptosis pathway in the SCI and Sham groups after treatment with Darapladib ( n = 3). Data are expressed as the mean ± SEM. * P < 0.05, ** P < 0.01, vs. SCI group (one-way analysis of variance followed by Tukey’s multiple comparison test). ASC: Apoptosis associated speck like protein containing CARD; CerS: ceramide synthase; DAPI: 4′,6-diamidino-2-phenylindole; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GSDMD: gasdermin D; GSDMD-N: N-term gasdermin D; IL: interleukin; NLRP3: NOD-like receptor thermal protein domain associated protein 3; Pla2g7: phospholipase a2 group VII; SCI: spinal cord injury; si.CerS5: CerS5 siRNA.
Techniques Used: Immunofluorescence, Staining, Western Blot, Expressing, Comparison


