Review



darapladib  (MedChemExpress)


Bioz Verified Symbol MedChemExpress is a verified supplier
Bioz Manufacturer Symbol MedChemExpress manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
    • 93
      Buy from Supplier

    Structured Review

    MedChemExpress darapladib
    A TRAP and F-actin staining in BMMs derived from Pla2g7 f/f and Pla2g7 LysM mice following osteoclast induction (scale bar, normal, 500 μm, enlarged, 200 μm). B Quantification of TRAP positive MNCs and average osteoclast size. n = 6 biologically independent wells per group. C , D Relative gene and protein expression of osteoclast markers, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. E The molecular structure diagram of <t>Darapladib.</t> F , G TRAP and F-actin staining and quantification of osteoclasts stimulated with RANKL in the absence or presence of Darapladib in a concentration-dependent manner (0, 100, 200, and 400 nM), with Darapladib administered throughout the entire induction period. n = 6 biologically independent wells per group. (scale bar, normal, 500 μm, enlarged, 200 μm). H , I The gene and protein expression of osteoclast markers. n = 3 biologically independent wells per group. J The immunoblotting analysis in osteoclasts induced by RANKL with or without Darapladib at different time points. K , L Hydroxyapatite resorption assay and statistical analysis of BMMs incubated with RANKL with or without Darapaldib (400 nmol/L). n = 3 biologically independent cell samples per group. (scale bar, normal, 200 μm). M TRAP and F-actin staining images of osteoclasts stimulated by RANKL in the absence or presence of recombinant Pla2g7 protein (50, 100, or 200 ng/mL) for 5 days, with Pla2g7 administered throughout the entire induction period (scale bar, normal, 500 μm, enlarged, 200 μm). N , O Relative gene and protein expression of osteoclast-specific markers. n = 3 biologically independent wells per group. P Western blot analysis of BMMs incubated with RANKL in the absence or presence of Pla2g7 at various time points. Data were showed as mean ± SD. Two-sided Student’s t test ( B – D , L ) was employed to assess the difference between the two groups. One-way ANOVA ( G ) and two-way ANOVA ( H , N ) were utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.
    Darapladib, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 93/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/darapladib/product/MedChemExpress
    Average 93 stars, based on 14 article reviews
    darapladib - by Bioz Stars, 2026-02
    93/100 stars

    Images

    1) Product Images from "Pla2g7 regulates bone homeostasis via Alox12/12-HETE/Gpr31 signaling axis"

    Article Title: Pla2g7 regulates bone homeostasis via Alox12/12-HETE/Gpr31 signaling axis

    Journal: Nature Communications

    doi: 10.1038/s41467-025-66285-8

    A TRAP and F-actin staining in BMMs derived from Pla2g7 f/f and Pla2g7 LysM mice following osteoclast induction (scale bar, normal, 500 μm, enlarged, 200 μm). B Quantification of TRAP positive MNCs and average osteoclast size. n = 6 biologically independent wells per group. C , D Relative gene and protein expression of osteoclast markers, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. E The molecular structure diagram of Darapladib. F , G TRAP and F-actin staining and quantification of osteoclasts stimulated with RANKL in the absence or presence of Darapladib in a concentration-dependent manner (0, 100, 200, and 400 nM), with Darapladib administered throughout the entire induction period. n = 6 biologically independent wells per group. (scale bar, normal, 500 μm, enlarged, 200 μm). H , I The gene and protein expression of osteoclast markers. n = 3 biologically independent wells per group. J The immunoblotting analysis in osteoclasts induced by RANKL with or without Darapladib at different time points. K , L Hydroxyapatite resorption assay and statistical analysis of BMMs incubated with RANKL with or without Darapaldib (400 nmol/L). n = 3 biologically independent cell samples per group. (scale bar, normal, 200 μm). M TRAP and F-actin staining images of osteoclasts stimulated by RANKL in the absence or presence of recombinant Pla2g7 protein (50, 100, or 200 ng/mL) for 5 days, with Pla2g7 administered throughout the entire induction period (scale bar, normal, 500 μm, enlarged, 200 μm). N , O Relative gene and protein expression of osteoclast-specific markers. n = 3 biologically independent wells per group. P Western blot analysis of BMMs incubated with RANKL in the absence or presence of Pla2g7 at various time points. Data were showed as mean ± SD. Two-sided Student’s t test ( B – D , L ) was employed to assess the difference between the two groups. One-way ANOVA ( G ) and two-way ANOVA ( H , N ) were utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.
    Figure Legend Snippet: A TRAP and F-actin staining in BMMs derived from Pla2g7 f/f and Pla2g7 LysM mice following osteoclast induction (scale bar, normal, 500 μm, enlarged, 200 μm). B Quantification of TRAP positive MNCs and average osteoclast size. n = 6 biologically independent wells per group. C , D Relative gene and protein expression of osteoclast markers, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. E The molecular structure diagram of Darapladib. F , G TRAP and F-actin staining and quantification of osteoclasts stimulated with RANKL in the absence or presence of Darapladib in a concentration-dependent manner (0, 100, 200, and 400 nM), with Darapladib administered throughout the entire induction period. n = 6 biologically independent wells per group. (scale bar, normal, 500 μm, enlarged, 200 μm). H , I The gene and protein expression of osteoclast markers. n = 3 biologically independent wells per group. J The immunoblotting analysis in osteoclasts induced by RANKL with or without Darapladib at different time points. K , L Hydroxyapatite resorption assay and statistical analysis of BMMs incubated with RANKL with or without Darapaldib (400 nmol/L). n = 3 biologically independent cell samples per group. (scale bar, normal, 200 μm). M TRAP and F-actin staining images of osteoclasts stimulated by RANKL in the absence or presence of recombinant Pla2g7 protein (50, 100, or 200 ng/mL) for 5 days, with Pla2g7 administered throughout the entire induction period (scale bar, normal, 500 μm, enlarged, 200 μm). N , O Relative gene and protein expression of osteoclast-specific markers. n = 3 biologically independent wells per group. P Western blot analysis of BMMs incubated with RANKL in the absence or presence of Pla2g7 at various time points. Data were showed as mean ± SD. Two-sided Student’s t test ( B – D , L ) was employed to assess the difference between the two groups. One-way ANOVA ( G ) and two-way ANOVA ( H , N ) were utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Techniques Used: Staining, Derivative Assay, Expressing, Western Blot, Concentration Assay, Incubation, Recombinant

    A The design of lipid metabolomics assay, in which Darapladib treatment was conducted concurrently with RANKL induction for 3 days until osteoclasts emerged. n = 5 biologically independent cell samples per group. B Volcano plot indicating differentially expressed genes in BMMs incubated with RANKL in the absence or presence of Darapladib (400 nmol/L). C Network analysis of the top 5 most significantly upregulated and downregulated secondary metabolites according to the integrated analysis of Fold change and P value. D Heatmap of differentially expressed secondary metabolites. FFA free fatty acid, SM sphingomyelin, LPC lysophosphatidylcholine, LPI lysophosphatidylinositol, LPE lysophosphatidylethanolamine, PC phosphatidylcholine, PS phosphatidylserine, PI phosphatidylinositol, PG phosphatidylglycerol, PE phosphatidylethanolamine. n = 5 biologically independent cell samples per group. E A schematic diagram of the RNA-sequencing of BMMs from WT mice with or without Darapladib, following 3 days of RANKL induction until osteoclasts emerged. n = 3 biologically independent cell samples per group. F Volcano plot showing 1036 upregulated and 1979 downregulated genes for RNA-seq data. G A heatmap of osteoclast-specific genes between RANKL and RANKL treated by Darapladib groups. n = 3 biologically independent cell samples per group. H The metabolism class in KEGG analysis of RNA-seq data. I , J The concentration of arachidonic acid in Darapladib, recombinant Pla2g7 and Pla2g7 cKO groups. n = 4 biologically independent mice per group. K – M The expression of ferroptosis-related proteins in Darapladib, recombinant Pla2g7 and Pla2g7 cKO groups, and these densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. Data were showed as mean ± SD. Two-sided Student’s t test ( I , J and K – M ) was employed to assess the difference between the two groups. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.
    Figure Legend Snippet: A The design of lipid metabolomics assay, in which Darapladib treatment was conducted concurrently with RANKL induction for 3 days until osteoclasts emerged. n = 5 biologically independent cell samples per group. B Volcano plot indicating differentially expressed genes in BMMs incubated with RANKL in the absence or presence of Darapladib (400 nmol/L). C Network analysis of the top 5 most significantly upregulated and downregulated secondary metabolites according to the integrated analysis of Fold change and P value. D Heatmap of differentially expressed secondary metabolites. FFA free fatty acid, SM sphingomyelin, LPC lysophosphatidylcholine, LPI lysophosphatidylinositol, LPE lysophosphatidylethanolamine, PC phosphatidylcholine, PS phosphatidylserine, PI phosphatidylinositol, PG phosphatidylglycerol, PE phosphatidylethanolamine. n = 5 biologically independent cell samples per group. E A schematic diagram of the RNA-sequencing of BMMs from WT mice with or without Darapladib, following 3 days of RANKL induction until osteoclasts emerged. n = 3 biologically independent cell samples per group. F Volcano plot showing 1036 upregulated and 1979 downregulated genes for RNA-seq data. G A heatmap of osteoclast-specific genes between RANKL and RANKL treated by Darapladib groups. n = 3 biologically independent cell samples per group. H The metabolism class in KEGG analysis of RNA-seq data. I , J The concentration of arachidonic acid in Darapladib, recombinant Pla2g7 and Pla2g7 cKO groups. n = 4 biologically independent mice per group. K – M The expression of ferroptosis-related proteins in Darapladib, recombinant Pla2g7 and Pla2g7 cKO groups, and these densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. Data were showed as mean ± SD. Two-sided Student’s t test ( I , J and K – M ) was employed to assess the difference between the two groups. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Techniques Used: Incubation, RNA Sequencing, Concentration Assay, Recombinant, Expressing, Western Blot

    A – D The expression of enzymes associated with the arachidonic acid metabolism pathway in osteoclasts treated with the inhibitor or ablation of Pla2g7 according to immunoblotting and relative RNA analysis. n = 3 biologically independent wells per group associated with cyclooxygenase (COX) and cytochrome P450 (CYP) pathways and n = 4 per group associated with lipoxygenase (LOX) pathways. E , F The immunofluorescence images of Alox12 in osteoclasts treated with Darapladib and quantification of fluorescence intensity. n = 3 biologically independent wells per group (scale bar, 10 μm). G , H The expression of Alox12 and Nfatc1 in mice treated with drug and statistical analysis of fluorescence intensity. n = 6 biologically independent mice per group (scale bar, 100 μm). I TRAP staining indicating Alox12 inhibitor treatment on Pla2g7-induced osteoclasts differentiation. (scale bar, 200 μm). J – L ML355 treatments (200 nM) on Pla2g7-induced osteoclasts in immunoblotting and RT-qPCR. n = 3 per group. n = 3 biologically independent wells per group. M The concentration of 12-HETE in RANKL or RANKL and Darapladib treatments were assessed following a 3-day induction period, coinciding with osteoclast formation. n = 4 biologically independent mice per group. N , O Darapladib or 12-HETE treatments (100 ng/mL) on RANKL-induced BMMs in TRAP staining and quantification of TRAP positive MNCs and average osteoclast size in each group. n = 6 biologically independent wells per group. P , Q Relative mRNA and protein expression in osteoclasts treated with Darapladib or 12-HETE, and the densitometric analysis of Western blot bands. For RNA analysis, n = 3 biologically independent wells per group. For Western blot analysis, n = 3 biologically independent cell samples per group. R Immunoblotting showing these markers in osteoclasts incubated with or without 12-HETE. Data were showed as mean ± SD. Two-sided Student’s t test ( B , D , F , H , M ) was employed to assess the difference between the two groups. Two-way ANOVA ( K , O – Q ) was utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.
    Figure Legend Snippet: A – D The expression of enzymes associated with the arachidonic acid metabolism pathway in osteoclasts treated with the inhibitor or ablation of Pla2g7 according to immunoblotting and relative RNA analysis. n = 3 biologically independent wells per group associated with cyclooxygenase (COX) and cytochrome P450 (CYP) pathways and n = 4 per group associated with lipoxygenase (LOX) pathways. E , F The immunofluorescence images of Alox12 in osteoclasts treated with Darapladib and quantification of fluorescence intensity. n = 3 biologically independent wells per group (scale bar, 10 μm). G , H The expression of Alox12 and Nfatc1 in mice treated with drug and statistical analysis of fluorescence intensity. n = 6 biologically independent mice per group (scale bar, 100 μm). I TRAP staining indicating Alox12 inhibitor treatment on Pla2g7-induced osteoclasts differentiation. (scale bar, 200 μm). J – L ML355 treatments (200 nM) on Pla2g7-induced osteoclasts in immunoblotting and RT-qPCR. n = 3 per group. n = 3 biologically independent wells per group. M The concentration of 12-HETE in RANKL or RANKL and Darapladib treatments were assessed following a 3-day induction period, coinciding with osteoclast formation. n = 4 biologically independent mice per group. N , O Darapladib or 12-HETE treatments (100 ng/mL) on RANKL-induced BMMs in TRAP staining and quantification of TRAP positive MNCs and average osteoclast size in each group. n = 6 biologically independent wells per group. P , Q Relative mRNA and protein expression in osteoclasts treated with Darapladib or 12-HETE, and the densitometric analysis of Western blot bands. For RNA analysis, n = 3 biologically independent wells per group. For Western blot analysis, n = 3 biologically independent cell samples per group. R Immunoblotting showing these markers in osteoclasts incubated with or without 12-HETE. Data were showed as mean ± SD. Two-sided Student’s t test ( B , D , F , H , M ) was employed to assess the difference between the two groups. Two-way ANOVA ( K , O – Q ) was utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Techniques Used: Expressing, Western Blot, Immunofluorescence, Fluorescence, Staining, Quantitative RT-PCR, Concentration Assay, Incubation

    A , B Knockdown efficiency of Gpr31 with siRNA from the level of gene and protein. n = 3 biologically independent wells per group. C TRAP staining to demonstrated the knockdown efficiency of Gpr31. D Immunoblotting showing deceased osteoclast markers expression in Gpr31 siRNA knockdown compared to control group during osteoclast formation. E KEGG analysis according to the differentially expressed genes of RNA-seq. F GSEA of oxidative phosphorylation and MAPK signaling pathway between RANKL and RANKL with Darapladib treatment. G , H The expression of MAPK signaling pathway in RANKL-induce osteoclast formation treated with drug or Gpr31 siRNA knockdown in Western blot. I – L OCR (left) and ECAR (right) of RANKL-induced BMMs with or without Darapladib, and statistical analysis of related parameters including Maximal respiration, ATP-linked respiration and Maximal ECAR. n = 3 biologically independent wells per group. M – P OCR (left) and ECAR (right) of BMMs incubated with Gpr31 siRNA compared with control group, and quantification of parameters Maximal respiration and Maximal ECAR. n = 3 biologically independent wells per group. ( Q , R ) The expression of mitochondrial respiratory chain, electron transport chain and osteoclast markers in WT BMMs treated with Darapladib or Gpr31 siRNA compared with control group. S The densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. T , U Immunoblotting showing the expression of mitochondrial dynamics and osteoclast markers in WT BMMs treated with Darapladib or Gpr31 siRNA compared with control group. V The densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. W – Z The flow diagram of MitoSOX and quantification of fluorescence intensity of probe. n = 3 biologically independent wells per group. Data were showed as mean ± SD. Two-sided Student’s t test ( S , V , Z ) was employed to assess the difference between the two groups. One-way ANOVA ( A , X ) and two-way ANOVA ( J , L , N , P ) were utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.
    Figure Legend Snippet: A , B Knockdown efficiency of Gpr31 with siRNA from the level of gene and protein. n = 3 biologically independent wells per group. C TRAP staining to demonstrated the knockdown efficiency of Gpr31. D Immunoblotting showing deceased osteoclast markers expression in Gpr31 siRNA knockdown compared to control group during osteoclast formation. E KEGG analysis according to the differentially expressed genes of RNA-seq. F GSEA of oxidative phosphorylation and MAPK signaling pathway between RANKL and RANKL with Darapladib treatment. G , H The expression of MAPK signaling pathway in RANKL-induce osteoclast formation treated with drug or Gpr31 siRNA knockdown in Western blot. I – L OCR (left) and ECAR (right) of RANKL-induced BMMs with or without Darapladib, and statistical analysis of related parameters including Maximal respiration, ATP-linked respiration and Maximal ECAR. n = 3 biologically independent wells per group. M – P OCR (left) and ECAR (right) of BMMs incubated with Gpr31 siRNA compared with control group, and quantification of parameters Maximal respiration and Maximal ECAR. n = 3 biologically independent wells per group. ( Q , R ) The expression of mitochondrial respiratory chain, electron transport chain and osteoclast markers in WT BMMs treated with Darapladib or Gpr31 siRNA compared with control group. S The densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. T , U Immunoblotting showing the expression of mitochondrial dynamics and osteoclast markers in WT BMMs treated with Darapladib or Gpr31 siRNA compared with control group. V The densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. W – Z The flow diagram of MitoSOX and quantification of fluorescence intensity of probe. n = 3 biologically independent wells per group. Data were showed as mean ± SD. Two-sided Student’s t test ( S , V , Z ) was employed to assess the difference between the two groups. One-way ANOVA ( A , X ) and two-way ANOVA ( J , L , N , P ) were utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Techniques Used: Knockdown, Staining, Western Blot, Expressing, Control, RNA Sequencing, Phospho-proteomics, Incubation, Fluorescence

    A The schematic diagram of high fat diets induced bone loss model. B Micro CT images of trabecular in CD or HFD treated with or without Darapladib groups. CD Chow diets, HFD high fat diets (scale bar, 1 mm). C Analysis of bone mass parameters in the three groups. n = 6 biologically independent mice per group. D , E Staining and statistical analysis of osteoclasts presence. n = 6 biologically independent mice per group. CD group served as control, HFD group represents high-fat diet model, and HFD + Darapladib group represents treatment with Darapladib on HFD background (scale bar, normal, 200 μm. enlarged, 100 μm). F , G Micro CT images and statistical analysis of cortical thickness. n = 6 biologically independent mice per group. (scale bar, 1 mm). H The schematic diagram of the ovariectomy experiment. I , J The images and bone mass analysis of trabecular in female mice divided into sham, OVX, OVX supplemented with Darapladib groups. n = 6 biologically independent mice per group (scale bar, 1 mm). K , L The expression of osteoclasts in two staining assays and statistical analysis. n = 6 biologically independent mice per group (scale bar, normal, 200 μm, enlarged, 100 μm). M , N The diagram of cortical bone and morphological parameters of WT, OVX and Darapladib-treated OVX mice. n = 6 biologically independent mice per group (scale bar, 1 mm). Data were showed as mean ± SD. Two-way ANOVA ( C , E , G , J , L , N ) was utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.
    Figure Legend Snippet: A The schematic diagram of high fat diets induced bone loss model. B Micro CT images of trabecular in CD or HFD treated with or without Darapladib groups. CD Chow diets, HFD high fat diets (scale bar, 1 mm). C Analysis of bone mass parameters in the three groups. n = 6 biologically independent mice per group. D , E Staining and statistical analysis of osteoclasts presence. n = 6 biologically independent mice per group. CD group served as control, HFD group represents high-fat diet model, and HFD + Darapladib group represents treatment with Darapladib on HFD background (scale bar, normal, 200 μm. enlarged, 100 μm). F , G Micro CT images and statistical analysis of cortical thickness. n = 6 biologically independent mice per group. (scale bar, 1 mm). H The schematic diagram of the ovariectomy experiment. I , J The images and bone mass analysis of trabecular in female mice divided into sham, OVX, OVX supplemented with Darapladib groups. n = 6 biologically independent mice per group (scale bar, 1 mm). K , L The expression of osteoclasts in two staining assays and statistical analysis. n = 6 biologically independent mice per group (scale bar, normal, 200 μm, enlarged, 100 μm). M , N The diagram of cortical bone and morphological parameters of WT, OVX and Darapladib-treated OVX mice. n = 6 biologically independent mice per group (scale bar, 1 mm). Data were showed as mean ± SD. Two-way ANOVA ( C , E , G , J , L , N ) was utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Techniques Used: Micro-CT, Staining, Control, Expressing

    A , B Immunohistochemistry staining and statistical analysis of Pla2g7 in normal patients (NP) and osteoporosis patients (OP). n = 16 biologically independent patients per group (scale bar, normal, 200 μm. enlarged, 25 μm). C , D The immunofluorescence images of Pla2g7 and NFATc1 in NP and OP (scale bar, 25 μm), and the statistical analysis of Pla2g7 + NFATc1 + cells. n = 6 biologically independent patients per group. E , F Relative gene expression of osteoclasts in NP and OP, and protein expression with densitometric analysis of Western blot bands. RNA, n = 6 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. G The expression of Pla2g7 and osteoclast markers during human osteoclast differentiation. H TRAP staining indicating Darapladib treatments (400 nM) on RANKL-induced osteoclast differentiation. (scale bar, 100 μm). I , J The expression of protein and gene in osteoclast differentiation stimulated with RANKL or RANKL and Darapladib, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. K TRAP staining indicating Pla2g7 treatments (200 ng/mL) on RANKL-induced osteoclast differentiation. (scale bar, 100 μm). L , M Immunoblotting and RT-qPCR showing increased osteoclast markers expression in Pla2g7 treatment group compared to control group, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. N , O ALP, ARS and protein expression in human mesenchymal stem cells (MSCs) incubated with standard induction mode with a dose-dependent manner of Darapladib (scale bar, 4 mm). P Osteoblast-related staining indicating the differentiation and mineralization of human mesenchymal stem cells treated with various concentration of Pla2g7. (scale bar, 4 mm). Q Immunoblotting showing the expression of osteoblast markers in human MSCs incubated with dexamethasone, ascorbic acid, β-glycerophosphate and Pla2g7 with different concentration Data were showed as mean ± SD. Two-sided Student’s t test ( B , D , E , F , I , J , L , M ) was employed to assess the difference between the two groups. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.
    Figure Legend Snippet: A , B Immunohistochemistry staining and statistical analysis of Pla2g7 in normal patients (NP) and osteoporosis patients (OP). n = 16 biologically independent patients per group (scale bar, normal, 200 μm. enlarged, 25 μm). C , D The immunofluorescence images of Pla2g7 and NFATc1 in NP and OP (scale bar, 25 μm), and the statistical analysis of Pla2g7 + NFATc1 + cells. n = 6 biologically independent patients per group. E , F Relative gene expression of osteoclasts in NP and OP, and protein expression with densitometric analysis of Western blot bands. RNA, n = 6 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. G The expression of Pla2g7 and osteoclast markers during human osteoclast differentiation. H TRAP staining indicating Darapladib treatments (400 nM) on RANKL-induced osteoclast differentiation. (scale bar, 100 μm). I , J The expression of protein and gene in osteoclast differentiation stimulated with RANKL or RANKL and Darapladib, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. K TRAP staining indicating Pla2g7 treatments (200 ng/mL) on RANKL-induced osteoclast differentiation. (scale bar, 100 μm). L , M Immunoblotting and RT-qPCR showing increased osteoclast markers expression in Pla2g7 treatment group compared to control group, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. N , O ALP, ARS and protein expression in human mesenchymal stem cells (MSCs) incubated with standard induction mode with a dose-dependent manner of Darapladib (scale bar, 4 mm). P Osteoblast-related staining indicating the differentiation and mineralization of human mesenchymal stem cells treated with various concentration of Pla2g7. (scale bar, 4 mm). Q Immunoblotting showing the expression of osteoblast markers in human MSCs incubated with dexamethasone, ascorbic acid, β-glycerophosphate and Pla2g7 with different concentration Data were showed as mean ± SD. Two-sided Student’s t test ( B , D , E , F , I , J , L , M ) was employed to assess the difference between the two groups. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Techniques Used: Immunohistochemistry, Staining, Immunofluorescence, Gene Expression, Expressing, Western Blot, Quantitative RT-PCR, Control, Incubation, Concentration Assay



    Similar Products

    darapladib  (MedChemExpress)
    93
    MedChemExpress darapladib
    A TRAP and F-actin staining in BMMs derived from Pla2g7 f/f and Pla2g7 LysM mice following osteoclast induction (scale bar, normal, 500 μm, enlarged, 200 μm). B Quantification of TRAP positive MNCs and average osteoclast size. n = 6 biologically independent wells per group. C , D Relative gene and protein expression of osteoclast markers, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. E The molecular structure diagram of <t>Darapladib.</t> F , G TRAP and F-actin staining and quantification of osteoclasts stimulated with RANKL in the absence or presence of Darapladib in a concentration-dependent manner (0, 100, 200, and 400 nM), with Darapladib administered throughout the entire induction period. n = 6 biologically independent wells per group. (scale bar, normal, 500 μm, enlarged, 200 μm). H , I The gene and protein expression of osteoclast markers. n = 3 biologically independent wells per group. J The immunoblotting analysis in osteoclasts induced by RANKL with or without Darapladib at different time points. K , L Hydroxyapatite resorption assay and statistical analysis of BMMs incubated with RANKL with or without Darapaldib (400 nmol/L). n = 3 biologically independent cell samples per group. (scale bar, normal, 200 μm). M TRAP and F-actin staining images of osteoclasts stimulated by RANKL in the absence or presence of recombinant Pla2g7 protein (50, 100, or 200 ng/mL) for 5 days, with Pla2g7 administered throughout the entire induction period (scale bar, normal, 500 μm, enlarged, 200 μm). N , O Relative gene and protein expression of osteoclast-specific markers. n = 3 biologically independent wells per group. P Western blot analysis of BMMs incubated with RANKL in the absence or presence of Pla2g7 at various time points. Data were showed as mean ± SD. Two-sided Student’s t test ( B – D , L ) was employed to assess the difference between the two groups. One-way ANOVA ( G ) and two-way ANOVA ( H , N ) were utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.
    Darapladib, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/darapladib/product/MedChemExpress
    Average 93 stars, based on 1 article reviews
    darapladib - by Bioz Stars, 2026-02
    93/100 stars
    		  Buy from Supplier
    darapladib  (TargetMol)
    94
    TargetMol darapladib
    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 , <t>Pla2g7</t> ), 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.
    Darapladib, supplied by TargetMol, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/darapladib/product/TargetMol
    Average 94 stars, based on 1 article reviews
    darapladib - by Bioz Stars, 2026-02
    94/100 stars
    		  Buy from Supplier
    darapladib  (Selleck Chemicals)
    92
    Selleck Chemicals darapladib
    A. P. falciparum parasites were incubated with Erastin (cystine uptake inhibitor) and Pyrimethamine to assess the relative reduction in DNA after 72 hours. No cooperation between the drugs was observed. Suppl. Fig. 17 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Erastin. B. Erastin2 (inhibitor of cystine uptake) with Pyrimethamine. Suppl. Fig. 18 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Erastin2. C. L -Buthionine- S,R -sulfoximine (BSO, inhibitor of glutamate-cysteine ligase) with Pyrimethamine. Suppl. Fig. 19 displays the values obtained from each biological replicate, which were plotted individually for each concentration of BSO. D. <t>Darapladib</t> (inhibitor of Peroxiredoxin 6) with Pyrimethamine. Suppl. Fig. 20 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Darapladib. E. Conoidin A (inhibitor of Peroxiredoxin 2) with Pyrimethamine. Suppl. Fig. 21 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Conoidin A.
    Darapladib, supplied by Selleck Chemicals, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/darapladib/product/Selleck Chemicals
    Average 92 stars, based on 1 article reviews
    darapladib - by Bioz Stars, 2026-02
    92/100 stars
    		  Buy from Supplier
    darapladib  (Cayman Chemical)
    90
    Cayman Chemical darapladib
    A. P. falciparum parasites were incubated with Erastin (cystine uptake inhibitor) and Pyrimethamine to assess the relative reduction in DNA after 72 hours. No cooperation between the drugs was observed. Suppl. Fig. 17 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Erastin. B. Erastin2 (inhibitor of cystine uptake) with Pyrimethamine. Suppl. Fig. 18 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Erastin2. C. L -Buthionine- S,R -sulfoximine (BSO, inhibitor of glutamate-cysteine ligase) with Pyrimethamine. Suppl. Fig. 19 displays the values obtained from each biological replicate, which were plotted individually for each concentration of BSO. D. <t>Darapladib</t> (inhibitor of Peroxiredoxin 6) with Pyrimethamine. Suppl. Fig. 20 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Darapladib. E. Conoidin A (inhibitor of Peroxiredoxin 2) with Pyrimethamine. Suppl. Fig. 21 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Conoidin A.
    Darapladib, supplied by Cayman Chemical, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/darapladib/product/Cayman Chemical
    Average 90 stars, based on 1 article reviews
    darapladib - by Bioz Stars, 2026-02
    90/100 stars
    		  Buy from Supplier
    mycobacterium complete medium  (MedChemExpress)
    94
    MedChemExpress mycobacterium complete medium
    A. P. falciparum parasites were incubated with Erastin (cystine uptake inhibitor) and Pyrimethamine to assess the relative reduction in DNA after 72 hours. No cooperation between the drugs was observed. Suppl. Fig. 17 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Erastin. B. Erastin2 (inhibitor of cystine uptake) with Pyrimethamine. Suppl. Fig. 18 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Erastin2. C. L -Buthionine- S,R -sulfoximine (BSO, inhibitor of glutamate-cysteine ligase) with Pyrimethamine. Suppl. Fig. 19 displays the values obtained from each biological replicate, which were plotted individually for each concentration of BSO. D. <t>Darapladib</t> (inhibitor of Peroxiredoxin 6) with Pyrimethamine. Suppl. Fig. 20 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Darapladib. E. Conoidin A (inhibitor of Peroxiredoxin 2) with Pyrimethamine. Suppl. Fig. 21 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Conoidin A.
    Mycobacterium Complete Medium, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mycobacterium complete medium/product/MedChemExpress
    Average 94 stars, based on 1 article reviews
    mycobacterium complete medium - by Bioz Stars, 2026-02
    94/100 stars
    		  Buy from Supplier
    plac test  (MedChemExpress)
    94
    MedChemExpress plac test
    Plasma PLA 2 enzymes and inhibitors used in this work
    Plac Test, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plac test/product/MedChemExpress
    Average 94 stars, based on 1 article reviews
    plac test - by Bioz Stars, 2026-02
    94/100 stars
    		  Buy from Supplier

    Image Search Results


    A TRAP and F-actin staining in BMMs derived from Pla2g7 f/f and Pla2g7 LysM mice following osteoclast induction (scale bar, normal, 500 μm, enlarged, 200 μm). B Quantification of TRAP positive MNCs and average osteoclast size. n = 6 biologically independent wells per group. C , D Relative gene and protein expression of osteoclast markers, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. E The molecular structure diagram of Darapladib. F , G TRAP and F-actin staining and quantification of osteoclasts stimulated with RANKL in the absence or presence of Darapladib in a concentration-dependent manner (0, 100, 200, and 400 nM), with Darapladib administered throughout the entire induction period. n = 6 biologically independent wells per group. (scale bar, normal, 500 μm, enlarged, 200 μm). H , I The gene and protein expression of osteoclast markers. n = 3 biologically independent wells per group. J The immunoblotting analysis in osteoclasts induced by RANKL with or without Darapladib at different time points. K , L Hydroxyapatite resorption assay and statistical analysis of BMMs incubated with RANKL with or without Darapaldib (400 nmol/L). n = 3 biologically independent cell samples per group. (scale bar, normal, 200 μm). M TRAP and F-actin staining images of osteoclasts stimulated by RANKL in the absence or presence of recombinant Pla2g7 protein (50, 100, or 200 ng/mL) for 5 days, with Pla2g7 administered throughout the entire induction period (scale bar, normal, 500 μm, enlarged, 200 μm). N , O Relative gene and protein expression of osteoclast-specific markers. n = 3 biologically independent wells per group. P Western blot analysis of BMMs incubated with RANKL in the absence or presence of Pla2g7 at various time points. Data were showed as mean ± SD. Two-sided Student’s t test ( B – D , L ) was employed to assess the difference between the two groups. One-way ANOVA ( G ) and two-way ANOVA ( H , N ) were utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Journal: Nature Communications

    Article Title: Pla2g7 regulates bone homeostasis via Alox12/12-HETE/Gpr31 signaling axis

    doi: 10.1038/s41467-025-66285-8

    Figure Lengend Snippet: A TRAP and F-actin staining in BMMs derived from Pla2g7 f/f and Pla2g7 LysM mice following osteoclast induction (scale bar, normal, 500 μm, enlarged, 200 μm). B Quantification of TRAP positive MNCs and average osteoclast size. n = 6 biologically independent wells per group. C , D Relative gene and protein expression of osteoclast markers, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. E The molecular structure diagram of Darapladib. F , G TRAP and F-actin staining and quantification of osteoclasts stimulated with RANKL in the absence or presence of Darapladib in a concentration-dependent manner (0, 100, 200, and 400 nM), with Darapladib administered throughout the entire induction period. n = 6 biologically independent wells per group. (scale bar, normal, 500 μm, enlarged, 200 μm). H , I The gene and protein expression of osteoclast markers. n = 3 biologically independent wells per group. J The immunoblotting analysis in osteoclasts induced by RANKL with or without Darapladib at different time points. K , L Hydroxyapatite resorption assay and statistical analysis of BMMs incubated with RANKL with or without Darapaldib (400 nmol/L). n = 3 biologically independent cell samples per group. (scale bar, normal, 200 μm). M TRAP and F-actin staining images of osteoclasts stimulated by RANKL in the absence or presence of recombinant Pla2g7 protein (50, 100, or 200 ng/mL) for 5 days, with Pla2g7 administered throughout the entire induction period (scale bar, normal, 500 μm, enlarged, 200 μm). N , O Relative gene and protein expression of osteoclast-specific markers. n = 3 biologically independent wells per group. P Western blot analysis of BMMs incubated with RANKL in the absence or presence of Pla2g7 at various time points. Data were showed as mean ± SD. Two-sided Student’s t test ( B – D , L ) was employed to assess the difference between the two groups. One-way ANOVA ( G ) and two-way ANOVA ( H , N ) were utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Article Snippet: ML355 (Cat#HY-12341) and Darapladib (Cat#HY-10521) were purchased from MedChemExpress.

    Techniques: Staining, Derivative Assay, Expressing, Western Blot, Concentration Assay, Incubation, Recombinant

    A The design of lipid metabolomics assay, in which Darapladib treatment was conducted concurrently with RANKL induction for 3 days until osteoclasts emerged. n = 5 biologically independent cell samples per group. B Volcano plot indicating differentially expressed genes in BMMs incubated with RANKL in the absence or presence of Darapladib (400 nmol/L). C Network analysis of the top 5 most significantly upregulated and downregulated secondary metabolites according to the integrated analysis of Fold change and P value. D Heatmap of differentially expressed secondary metabolites. FFA free fatty acid, SM sphingomyelin, LPC lysophosphatidylcholine, LPI lysophosphatidylinositol, LPE lysophosphatidylethanolamine, PC phosphatidylcholine, PS phosphatidylserine, PI phosphatidylinositol, PG phosphatidylglycerol, PE phosphatidylethanolamine. n = 5 biologically independent cell samples per group. E A schematic diagram of the RNA-sequencing of BMMs from WT mice with or without Darapladib, following 3 days of RANKL induction until osteoclasts emerged. n = 3 biologically independent cell samples per group. F Volcano plot showing 1036 upregulated and 1979 downregulated genes for RNA-seq data. G A heatmap of osteoclast-specific genes between RANKL and RANKL treated by Darapladib groups. n = 3 biologically independent cell samples per group. H The metabolism class in KEGG analysis of RNA-seq data. I , J The concentration of arachidonic acid in Darapladib, recombinant Pla2g7 and Pla2g7 cKO groups. n = 4 biologically independent mice per group. K – M The expression of ferroptosis-related proteins in Darapladib, recombinant Pla2g7 and Pla2g7 cKO groups, and these densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. Data were showed as mean ± SD. Two-sided Student’s t test ( I , J and K – M ) was employed to assess the difference between the two groups. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Journal: Nature Communications

    Article Title: Pla2g7 regulates bone homeostasis via Alox12/12-HETE/Gpr31 signaling axis

    doi: 10.1038/s41467-025-66285-8

    Figure Lengend Snippet: A The design of lipid metabolomics assay, in which Darapladib treatment was conducted concurrently with RANKL induction for 3 days until osteoclasts emerged. n = 5 biologically independent cell samples per group. B Volcano plot indicating differentially expressed genes in BMMs incubated with RANKL in the absence or presence of Darapladib (400 nmol/L). C Network analysis of the top 5 most significantly upregulated and downregulated secondary metabolites according to the integrated analysis of Fold change and P value. D Heatmap of differentially expressed secondary metabolites. FFA free fatty acid, SM sphingomyelin, LPC lysophosphatidylcholine, LPI lysophosphatidylinositol, LPE lysophosphatidylethanolamine, PC phosphatidylcholine, PS phosphatidylserine, PI phosphatidylinositol, PG phosphatidylglycerol, PE phosphatidylethanolamine. n = 5 biologically independent cell samples per group. E A schematic diagram of the RNA-sequencing of BMMs from WT mice with or without Darapladib, following 3 days of RANKL induction until osteoclasts emerged. n = 3 biologically independent cell samples per group. F Volcano plot showing 1036 upregulated and 1979 downregulated genes for RNA-seq data. G A heatmap of osteoclast-specific genes between RANKL and RANKL treated by Darapladib groups. n = 3 biologically independent cell samples per group. H The metabolism class in KEGG analysis of RNA-seq data. I , J The concentration of arachidonic acid in Darapladib, recombinant Pla2g7 and Pla2g7 cKO groups. n = 4 biologically independent mice per group. K – M The expression of ferroptosis-related proteins in Darapladib, recombinant Pla2g7 and Pla2g7 cKO groups, and these densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. Data were showed as mean ± SD. Two-sided Student’s t test ( I , J and K – M ) was employed to assess the difference between the two groups. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Article Snippet: ML355 (Cat#HY-12341) and Darapladib (Cat#HY-10521) were purchased from MedChemExpress.

    Techniques: Incubation, RNA Sequencing, Concentration Assay, Recombinant, Expressing, Western Blot

    A – D The expression of enzymes associated with the arachidonic acid metabolism pathway in osteoclasts treated with the inhibitor or ablation of Pla2g7 according to immunoblotting and relative RNA analysis. n = 3 biologically independent wells per group associated with cyclooxygenase (COX) and cytochrome P450 (CYP) pathways and n = 4 per group associated with lipoxygenase (LOX) pathways. E , F The immunofluorescence images of Alox12 in osteoclasts treated with Darapladib and quantification of fluorescence intensity. n = 3 biologically independent wells per group (scale bar, 10 μm). G , H The expression of Alox12 and Nfatc1 in mice treated with drug and statistical analysis of fluorescence intensity. n = 6 biologically independent mice per group (scale bar, 100 μm). I TRAP staining indicating Alox12 inhibitor treatment on Pla2g7-induced osteoclasts differentiation. (scale bar, 200 μm). J – L ML355 treatments (200 nM) on Pla2g7-induced osteoclasts in immunoblotting and RT-qPCR. n = 3 per group. n = 3 biologically independent wells per group. M The concentration of 12-HETE in RANKL or RANKL and Darapladib treatments were assessed following a 3-day induction period, coinciding with osteoclast formation. n = 4 biologically independent mice per group. N , O Darapladib or 12-HETE treatments (100 ng/mL) on RANKL-induced BMMs in TRAP staining and quantification of TRAP positive MNCs and average osteoclast size in each group. n = 6 biologically independent wells per group. P , Q Relative mRNA and protein expression in osteoclasts treated with Darapladib or 12-HETE, and the densitometric analysis of Western blot bands. For RNA analysis, n = 3 biologically independent wells per group. For Western blot analysis, n = 3 biologically independent cell samples per group. R Immunoblotting showing these markers in osteoclasts incubated with or without 12-HETE. Data were showed as mean ± SD. Two-sided Student’s t test ( B , D , F , H , M ) was employed to assess the difference between the two groups. Two-way ANOVA ( K , O – Q ) was utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Journal: Nature Communications

    Article Title: Pla2g7 regulates bone homeostasis via Alox12/12-HETE/Gpr31 signaling axis

    doi: 10.1038/s41467-025-66285-8

    Figure Lengend Snippet: A – D The expression of enzymes associated with the arachidonic acid metabolism pathway in osteoclasts treated with the inhibitor or ablation of Pla2g7 according to immunoblotting and relative RNA analysis. n = 3 biologically independent wells per group associated with cyclooxygenase (COX) and cytochrome P450 (CYP) pathways and n = 4 per group associated with lipoxygenase (LOX) pathways. E , F The immunofluorescence images of Alox12 in osteoclasts treated with Darapladib and quantification of fluorescence intensity. n = 3 biologically independent wells per group (scale bar, 10 μm). G , H The expression of Alox12 and Nfatc1 in mice treated with drug and statistical analysis of fluorescence intensity. n = 6 biologically independent mice per group (scale bar, 100 μm). I TRAP staining indicating Alox12 inhibitor treatment on Pla2g7-induced osteoclasts differentiation. (scale bar, 200 μm). J – L ML355 treatments (200 nM) on Pla2g7-induced osteoclasts in immunoblotting and RT-qPCR. n = 3 per group. n = 3 biologically independent wells per group. M The concentration of 12-HETE in RANKL or RANKL and Darapladib treatments were assessed following a 3-day induction period, coinciding with osteoclast formation. n = 4 biologically independent mice per group. N , O Darapladib or 12-HETE treatments (100 ng/mL) on RANKL-induced BMMs in TRAP staining and quantification of TRAP positive MNCs and average osteoclast size in each group. n = 6 biologically independent wells per group. P , Q Relative mRNA and protein expression in osteoclasts treated with Darapladib or 12-HETE, and the densitometric analysis of Western blot bands. For RNA analysis, n = 3 biologically independent wells per group. For Western blot analysis, n = 3 biologically independent cell samples per group. R Immunoblotting showing these markers in osteoclasts incubated with or without 12-HETE. Data were showed as mean ± SD. Two-sided Student’s t test ( B , D , F , H , M ) was employed to assess the difference between the two groups. Two-way ANOVA ( K , O – Q ) was utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Article Snippet: ML355 (Cat#HY-12341) and Darapladib (Cat#HY-10521) were purchased from MedChemExpress.

    Techniques: Expressing, Western Blot, Immunofluorescence, Fluorescence, Staining, Quantitative RT-PCR, Concentration Assay, Incubation

    A , B Knockdown efficiency of Gpr31 with siRNA from the level of gene and protein. n = 3 biologically independent wells per group. C TRAP staining to demonstrated the knockdown efficiency of Gpr31. D Immunoblotting showing deceased osteoclast markers expression in Gpr31 siRNA knockdown compared to control group during osteoclast formation. E KEGG analysis according to the differentially expressed genes of RNA-seq. F GSEA of oxidative phosphorylation and MAPK signaling pathway between RANKL and RANKL with Darapladib treatment. G , H The expression of MAPK signaling pathway in RANKL-induce osteoclast formation treated with drug or Gpr31 siRNA knockdown in Western blot. I – L OCR (left) and ECAR (right) of RANKL-induced BMMs with or without Darapladib, and statistical analysis of related parameters including Maximal respiration, ATP-linked respiration and Maximal ECAR. n = 3 biologically independent wells per group. M – P OCR (left) and ECAR (right) of BMMs incubated with Gpr31 siRNA compared with control group, and quantification of parameters Maximal respiration and Maximal ECAR. n = 3 biologically independent wells per group. ( Q , R ) The expression of mitochondrial respiratory chain, electron transport chain and osteoclast markers in WT BMMs treated with Darapladib or Gpr31 siRNA compared with control group. S The densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. T , U Immunoblotting showing the expression of mitochondrial dynamics and osteoclast markers in WT BMMs treated with Darapladib or Gpr31 siRNA compared with control group. V The densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. W – Z The flow diagram of MitoSOX and quantification of fluorescence intensity of probe. n = 3 biologically independent wells per group. Data were showed as mean ± SD. Two-sided Student’s t test ( S , V , Z ) was employed to assess the difference between the two groups. One-way ANOVA ( A , X ) and two-way ANOVA ( J , L , N , P ) were utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Journal: Nature Communications

    Article Title: Pla2g7 regulates bone homeostasis via Alox12/12-HETE/Gpr31 signaling axis

    doi: 10.1038/s41467-025-66285-8

    Figure Lengend Snippet: A , B Knockdown efficiency of Gpr31 with siRNA from the level of gene and protein. n = 3 biologically independent wells per group. C TRAP staining to demonstrated the knockdown efficiency of Gpr31. D Immunoblotting showing deceased osteoclast markers expression in Gpr31 siRNA knockdown compared to control group during osteoclast formation. E KEGG analysis according to the differentially expressed genes of RNA-seq. F GSEA of oxidative phosphorylation and MAPK signaling pathway between RANKL and RANKL with Darapladib treatment. G , H The expression of MAPK signaling pathway in RANKL-induce osteoclast formation treated with drug or Gpr31 siRNA knockdown in Western blot. I – L OCR (left) and ECAR (right) of RANKL-induced BMMs with or without Darapladib, and statistical analysis of related parameters including Maximal respiration, ATP-linked respiration and Maximal ECAR. n = 3 biologically independent wells per group. M – P OCR (left) and ECAR (right) of BMMs incubated with Gpr31 siRNA compared with control group, and quantification of parameters Maximal respiration and Maximal ECAR. n = 3 biologically independent wells per group. ( Q , R ) The expression of mitochondrial respiratory chain, electron transport chain and osteoclast markers in WT BMMs treated with Darapladib or Gpr31 siRNA compared with control group. S The densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. T , U Immunoblotting showing the expression of mitochondrial dynamics and osteoclast markers in WT BMMs treated with Darapladib or Gpr31 siRNA compared with control group. V The densitometric analysis of Western blot bands. n = 3 biologically independent cell samples per group. W – Z The flow diagram of MitoSOX and quantification of fluorescence intensity of probe. n = 3 biologically independent wells per group. Data were showed as mean ± SD. Two-sided Student’s t test ( S , V , Z ) was employed to assess the difference between the two groups. One-way ANOVA ( A , X ) and two-way ANOVA ( J , L , N , P ) were utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Article Snippet: ML355 (Cat#HY-12341) and Darapladib (Cat#HY-10521) were purchased from MedChemExpress.

    Techniques: Knockdown, Staining, Western Blot, Expressing, Control, RNA Sequencing, Phospho-proteomics, Incubation, Fluorescence

    A The schematic diagram of high fat diets induced bone loss model. B Micro CT images of trabecular in CD or HFD treated with or without Darapladib groups. CD Chow diets, HFD high fat diets (scale bar, 1 mm). C Analysis of bone mass parameters in the three groups. n = 6 biologically independent mice per group. D , E Staining and statistical analysis of osteoclasts presence. n = 6 biologically independent mice per group. CD group served as control, HFD group represents high-fat diet model, and HFD + Darapladib group represents treatment with Darapladib on HFD background (scale bar, normal, 200 μm. enlarged, 100 μm). F , G Micro CT images and statistical analysis of cortical thickness. n = 6 biologically independent mice per group. (scale bar, 1 mm). H The schematic diagram of the ovariectomy experiment. I , J The images and bone mass analysis of trabecular in female mice divided into sham, OVX, OVX supplemented with Darapladib groups. n = 6 biologically independent mice per group (scale bar, 1 mm). K , L The expression of osteoclasts in two staining assays and statistical analysis. n = 6 biologically independent mice per group (scale bar, normal, 200 μm, enlarged, 100 μm). M , N The diagram of cortical bone and morphological parameters of WT, OVX and Darapladib-treated OVX mice. n = 6 biologically independent mice per group (scale bar, 1 mm). Data were showed as mean ± SD. Two-way ANOVA ( C , E , G , J , L , N ) was utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Journal: Nature Communications

    Article Title: Pla2g7 regulates bone homeostasis via Alox12/12-HETE/Gpr31 signaling axis

    doi: 10.1038/s41467-025-66285-8

    Figure Lengend Snippet: A The schematic diagram of high fat diets induced bone loss model. B Micro CT images of trabecular in CD or HFD treated with or without Darapladib groups. CD Chow diets, HFD high fat diets (scale bar, 1 mm). C Analysis of bone mass parameters in the three groups. n = 6 biologically independent mice per group. D , E Staining and statistical analysis of osteoclasts presence. n = 6 biologically independent mice per group. CD group served as control, HFD group represents high-fat diet model, and HFD + Darapladib group represents treatment with Darapladib on HFD background (scale bar, normal, 200 μm. enlarged, 100 μm). F , G Micro CT images and statistical analysis of cortical thickness. n = 6 biologically independent mice per group. (scale bar, 1 mm). H The schematic diagram of the ovariectomy experiment. I , J The images and bone mass analysis of trabecular in female mice divided into sham, OVX, OVX supplemented with Darapladib groups. n = 6 biologically independent mice per group (scale bar, 1 mm). K , L The expression of osteoclasts in two staining assays and statistical analysis. n = 6 biologically independent mice per group (scale bar, normal, 200 μm, enlarged, 100 μm). M , N The diagram of cortical bone and morphological parameters of WT, OVX and Darapladib-treated OVX mice. n = 6 biologically independent mice per group (scale bar, 1 mm). Data were showed as mean ± SD. Two-way ANOVA ( C , E , G , J , L , N ) was utilized for multiple comparisons test. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Article Snippet: ML355 (Cat#HY-12341) and Darapladib (Cat#HY-10521) were purchased from MedChemExpress.

    Techniques: Micro-CT, Staining, Control, Expressing

    A , B Immunohistochemistry staining and statistical analysis of Pla2g7 in normal patients (NP) and osteoporosis patients (OP). n = 16 biologically independent patients per group (scale bar, normal, 200 μm. enlarged, 25 μm). C , D The immunofluorescence images of Pla2g7 and NFATc1 in NP and OP (scale bar, 25 μm), and the statistical analysis of Pla2g7 + NFATc1 + cells. n = 6 biologically independent patients per group. E , F Relative gene expression of osteoclasts in NP and OP, and protein expression with densitometric analysis of Western blot bands. RNA, n = 6 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. G The expression of Pla2g7 and osteoclast markers during human osteoclast differentiation. H TRAP staining indicating Darapladib treatments (400 nM) on RANKL-induced osteoclast differentiation. (scale bar, 100 μm). I , J The expression of protein and gene in osteoclast differentiation stimulated with RANKL or RANKL and Darapladib, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. K TRAP staining indicating Pla2g7 treatments (200 ng/mL) on RANKL-induced osteoclast differentiation. (scale bar, 100 μm). L , M Immunoblotting and RT-qPCR showing increased osteoclast markers expression in Pla2g7 treatment group compared to control group, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. N , O ALP, ARS and protein expression in human mesenchymal stem cells (MSCs) incubated with standard induction mode with a dose-dependent manner of Darapladib (scale bar, 4 mm). P Osteoblast-related staining indicating the differentiation and mineralization of human mesenchymal stem cells treated with various concentration of Pla2g7. (scale bar, 4 mm). Q Immunoblotting showing the expression of osteoblast markers in human MSCs incubated with dexamethasone, ascorbic acid, β-glycerophosphate and Pla2g7 with different concentration Data were showed as mean ± SD. Two-sided Student’s t test ( B , D , E , F , I , J , L , M ) was employed to assess the difference between the two groups. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Journal: Nature Communications

    Article Title: Pla2g7 regulates bone homeostasis via Alox12/12-HETE/Gpr31 signaling axis

    doi: 10.1038/s41467-025-66285-8

    Figure Lengend Snippet: A , B Immunohistochemistry staining and statistical analysis of Pla2g7 in normal patients (NP) and osteoporosis patients (OP). n = 16 biologically independent patients per group (scale bar, normal, 200 μm. enlarged, 25 μm). C , D The immunofluorescence images of Pla2g7 and NFATc1 in NP and OP (scale bar, 25 μm), and the statistical analysis of Pla2g7 + NFATc1 + cells. n = 6 biologically independent patients per group. E , F Relative gene expression of osteoclasts in NP and OP, and protein expression with densitometric analysis of Western blot bands. RNA, n = 6 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. G The expression of Pla2g7 and osteoclast markers during human osteoclast differentiation. H TRAP staining indicating Darapladib treatments (400 nM) on RANKL-induced osteoclast differentiation. (scale bar, 100 μm). I , J The expression of protein and gene in osteoclast differentiation stimulated with RANKL or RANKL and Darapladib, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. K TRAP staining indicating Pla2g7 treatments (200 ng/mL) on RANKL-induced osteoclast differentiation. (scale bar, 100 μm). L , M Immunoblotting and RT-qPCR showing increased osteoclast markers expression in Pla2g7 treatment group compared to control group, and the densitometric analysis of Western blot bands. RNA, n = 3 biologically independent wells per group. WB, n = 3 biologically independent cell samples per group. N , O ALP, ARS and protein expression in human mesenchymal stem cells (MSCs) incubated with standard induction mode with a dose-dependent manner of Darapladib (scale bar, 4 mm). P Osteoblast-related staining indicating the differentiation and mineralization of human mesenchymal stem cells treated with various concentration of Pla2g7. (scale bar, 4 mm). Q Immunoblotting showing the expression of osteoblast markers in human MSCs incubated with dexamethasone, ascorbic acid, β-glycerophosphate and Pla2g7 with different concentration Data were showed as mean ± SD. Two-sided Student’s t test ( B , D , E , F , I , J , L , M ) was employed to assess the difference between the two groups. Significance: * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001, ns non-significance.

    Article Snippet: ML355 (Cat#HY-12341) and Darapladib (Cat#HY-10521) were purchased from MedChemExpress.

    Techniques: Immunohistochemistry, Staining, Immunofluorescence, Gene Expression, Expressing, Western Blot, Quantitative RT-PCR, Control, Incubation, Concentration Assay

    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.

    Journal: Neural Regeneration Research

    Article Title: Inhibiting ceramide synthase 5 expression in microglia decreases neuroinflammation after spinal cord injury

    doi: 10.4103/NRR.NRR-D-23-01933

    Figure Lengend 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.

    Article Snippet: The mice were randomly assigned to one of five groups (10 mice per group): (1) Sham group, with no SCI induced after opening the vertebral plate; (2) SCI group, with no intrathecal injection after striking the spinal cord; (3) SCI + si.CerS5 group, which were injected with 10 nmol CerS5 siRNA (si.CerS5) (synthesized by GenePharm, Shanghai, China; ) intrathecally around the notch immediately after contusion; (4) SCI + si.NC group, which were injected intrathecally with si.NC (10 nmol) (synthesized by GenePharm, Shanghai, China; ) around the notch immediately after contusion; (5) SCI + Darapladib group, which were injected intrathecally with Darapladib (a Pla2g7 inhibitor, 10 mg/kg, TargetMol, T6109, Boston, MA, USA) around the notch immediately after contusion; and (6) SCI + Vehicle group, which were injected intrathecally with vehicle (10% dimethylsulfoxide, 40% polyethylene glycol 300, 5% Tween-80, and 45% phosphate-buffered saline) around the notch immediately after contusion.

    Techniques: Single-cell Analysis, Marker, Expressing

    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.

    Journal: Neural Regeneration Research

    Article Title: Inhibiting ceramide synthase 5 expression in microglia decreases neuroinflammation after spinal cord injury

    doi: 10.4103/NRR.NRR-D-23-01933

    Figure Lengend 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.

    Article Snippet: The mice were randomly assigned to one of five groups (10 mice per group): (1) Sham group, with no SCI induced after opening the vertebral plate; (2) SCI group, with no intrathecal injection after striking the spinal cord; (3) SCI + si.CerS5 group, which were injected with 10 nmol CerS5 siRNA (si.CerS5) (synthesized by GenePharm, Shanghai, China; ) intrathecally around the notch immediately after contusion; (4) SCI + si.NC group, which were injected intrathecally with si.NC (10 nmol) (synthesized by GenePharm, Shanghai, China; ) around the notch immediately after contusion; (5) SCI + Darapladib group, which were injected intrathecally with Darapladib (a Pla2g7 inhibitor, 10 mg/kg, TargetMol, T6109, Boston, MA, USA) around the notch immediately after contusion; and (6) SCI + Vehicle group, which were injected intrathecally with vehicle (10% dimethylsulfoxide, 40% polyethylene glycol 300, 5% Tween-80, and 45% phosphate-buffered saline) around the notch immediately after contusion.

    Techniques: Knockdown, Expressing, Western Blot, Immunofluorescence, Staining, Comparison, Binding Assay

    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.

    Journal: Neural Regeneration Research

    Article Title: Inhibiting ceramide synthase 5 expression in microglia decreases neuroinflammation after spinal cord injury

    doi: 10.4103/NRR.NRR-D-23-01933

    Figure Lengend 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.

    Article Snippet: The mice were randomly assigned to one of five groups (10 mice per group): (1) Sham group, with no SCI induced after opening the vertebral plate; (2) SCI group, with no intrathecal injection after striking the spinal cord; (3) SCI + si.CerS5 group, which were injected with 10 nmol CerS5 siRNA (si.CerS5) (synthesized by GenePharm, Shanghai, China; ) intrathecally around the notch immediately after contusion; (4) SCI + si.NC group, which were injected intrathecally with si.NC (10 nmol) (synthesized by GenePharm, Shanghai, China; ) around the notch immediately after contusion; (5) SCI + Darapladib group, which were injected intrathecally with Darapladib (a Pla2g7 inhibitor, 10 mg/kg, TargetMol, T6109, Boston, MA, USA) around the notch immediately after contusion; and (6) SCI + Vehicle group, which were injected intrathecally with vehicle (10% dimethylsulfoxide, 40% polyethylene glycol 300, 5% Tween-80, and 45% phosphate-buffered saline) around the notch immediately after contusion.

    Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Immunofluorescence, Staining, Comparison, Binding Assay

    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.

    Journal: Neural Regeneration Research

    Article Title: Inhibiting ceramide synthase 5 expression in microglia decreases neuroinflammation after spinal cord injury

    doi: 10.4103/NRR.NRR-D-23-01933

    Figure Lengend 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.

    Article Snippet: The mice were randomly assigned to one of five groups (10 mice per group): (1) Sham group, with no SCI induced after opening the vertebral plate; (2) SCI group, with no intrathecal injection after striking the spinal cord; (3) SCI + si.CerS5 group, which were injected with 10 nmol CerS5 siRNA (si.CerS5) (synthesized by GenePharm, Shanghai, China; ) intrathecally around the notch immediately after contusion; (4) SCI + si.NC group, which were injected intrathecally with si.NC (10 nmol) (synthesized by GenePharm, Shanghai, China; ) around the notch immediately after contusion; (5) SCI + Darapladib group, which were injected intrathecally with Darapladib (a Pla2g7 inhibitor, 10 mg/kg, TargetMol, T6109, Boston, MA, USA) around the notch immediately after contusion; and (6) SCI + Vehicle group, which were injected intrathecally with vehicle (10% dimethylsulfoxide, 40% polyethylene glycol 300, 5% Tween-80, and 45% phosphate-buffered saline) around the notch immediately after contusion.

    Techniques: Immunofluorescence, Staining, Western Blot, Expressing, Comparison

    A. P. falciparum parasites were incubated with Erastin (cystine uptake inhibitor) and Pyrimethamine to assess the relative reduction in DNA after 72 hours. No cooperation between the drugs was observed. Suppl. Fig. 17 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Erastin. B. Erastin2 (inhibitor of cystine uptake) with Pyrimethamine. Suppl. Fig. 18 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Erastin2. C. L -Buthionine- S,R -sulfoximine (BSO, inhibitor of glutamate-cysteine ligase) with Pyrimethamine. Suppl. Fig. 19 displays the values obtained from each biological replicate, which were plotted individually for each concentration of BSO. D. Darapladib (inhibitor of Peroxiredoxin 6) with Pyrimethamine. Suppl. Fig. 20 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Darapladib. E. Conoidin A (inhibitor of Peroxiredoxin 2) with Pyrimethamine. Suppl. Fig. 21 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Conoidin A.

    Journal: bioRxiv

    Article Title: Synergistic activity of the ferroptosis inducer RSL3 and Pyrimethamine to inhibit the proliferation of Plasmodium falciparum

    doi: 10.1101/2025.03.14.643256

    Figure Lengend Snippet: A. P. falciparum parasites were incubated with Erastin (cystine uptake inhibitor) and Pyrimethamine to assess the relative reduction in DNA after 72 hours. No cooperation between the drugs was observed. Suppl. Fig. 17 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Erastin. B. Erastin2 (inhibitor of cystine uptake) with Pyrimethamine. Suppl. Fig. 18 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Erastin2. C. L -Buthionine- S,R -sulfoximine (BSO, inhibitor of glutamate-cysteine ligase) with Pyrimethamine. Suppl. Fig. 19 displays the values obtained from each biological replicate, which were plotted individually for each concentration of BSO. D. Darapladib (inhibitor of Peroxiredoxin 6) with Pyrimethamine. Suppl. Fig. 20 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Darapladib. E. Conoidin A (inhibitor of Peroxiredoxin 2) with Pyrimethamine. Suppl. Fig. 21 displays the values obtained from each biological replicate, which were plotted individually for each concentration of Conoidin A.

    Article Snippet: Dihydroartemisinin, Atovaquone, Pyrimethamine, Darapladib, were provided by Selleckchem.

    Techniques: Incubation, Concentration Assay

    Plasma PLA 2 enzymes and inhibitors used in this work

    Journal: Journal of Lipid Research

    Article Title: Contribution of individual phospholipase A 2 enzymes to the cleavage of oxidized phospholipids in human blood plasma

    doi: 10.1016/j.jlr.2025.100742

    Figure Lengend Snippet: Plasma PLA 2 enzymes and inhibitors used in this work

    Article Snippet: Q13093 , PLA2G7 PAF-AH , 180 , 250 , PLAC test ( ) , Darapladib , 10 nM , www.medchemexpress.com/Darapladib.

    Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay