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Cayman Chemical gsk199

HFFs were infected with HSV-1 (MOI 1) and then treated with increasing concentrations of HF4 (A) or CAY10727 (B) , two PAD3-specific inhibitors, which was given 1 h prior to virus adsorption and kept throughout the whole experiment. At 24 hpi, viral plaques were microscopically counted, and the number of plaques was plotted as a function of inhibitor concentration. Values are expressed as means ± SEM (error bars) of three independent experiments. Values are expressed as mean ± SEM of three independent experiments, * P < 0.05, ** P < 0.01, *** P < 0.001; one-way ANOVA followed by Bonferroni’s post test. ( C ) Representative images of infected HFFs (24 hpi) at an MOI of 1 PFU/cell and treated with HF4 (5 μM), CAY10727 (1 μM), or vehicle (DMSO). ( D ) Protein lysates from uninfected (mock) or infected HFFs (24 hpi) at an MOI of 1 PFU/cell treated with AFM30a (20 μM), HF4 (5 μM), <t>GSK199</t> (20 μM) or vehicle (DMSO) were analyzed by immunoblotting to assess for viral expression with an anti-gD antibody; β-actin cellular expression was used for protein loading control. SH-SY5Y, ARPE-19, and HEK293 cells were infected with HSV-1 (MOI 1 PFU/cell) and then treated with increasing concentrations of HF4 (E) or CAY10727 (F) , which were given 1 h prior to virus adsorption and kept throughout the whole experiment. At 24 hpi, viral plaques were microscopically counted, and the number of plaques was plotted as a function of inhibitor concentration. Values are expressed as means ± SEM (error bars) of four independent experiments, * P < 0.05, ** P < 0.01, *** P < 0.001; one-way ANOVA followed by Bonferroni’s post test. (G) HFFs were silenced for PAD3 using specific siRNAs (siPAD3), as negative control cells were also similarly transfected with scrambled siRNA (siCTRL). At 24 h post-treatment (hpt), cells were infected with HSV-1 at an MOI of 1 PFU/cell. The efficiency of PAD3 protein depletion at 24 hpi was assessed by immunoblotting using antibodies against PAD3 or β-actin for equal loading. An anti-gD antibody was used to verify HSV-1 infection. Representative blots of three independent experiments are shown. ( H ) PAD3-silenced cells were infected with HSV-1 at an MOI of 1 PFU/cell. Viral supernatants were collected at 24 hpi and analyzed by standard plaque assay. Values are expressed as means ± SEM. Values are expressed as mean ± SEM of three independent experiments, *** P < 0.001; one-way ANOVA followed by Bonferroni’s post test. ( I ) Representative images of infected HFFs (24 hpi) at an MOI of 1 PFU/cell and transfected with the same siCTRL and siPAD3 described in the legend to .

Gsk199, 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/product/gsk199/gsk199/pmc10727434-165-7-16
Average 90 stars, based on 1 article reviews

gsk199 - by Bioz Stars, 2026-09

90/100 stars

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1) Product Images from "Citrullination profile analysis reveals peptidylarginine deaminase 3 as an HSV-1 target to dampen the activity of candidate antiviral restriction factors"

Article Title: Citrullination profile analysis reveals peptidylarginine deaminase 3 as an HSV-1 target to dampen the activity of candidate antiviral restriction factors

Journal: PLOS Pathogens

doi: 10.1371/journal.ppat.1011849

Figure Legend Snippet: HFFs were infected with HSV-1 (MOI 1) and then treated with increasing concentrations of HF4 (A) or CAY10727 (B) , two PAD3-specific inhibitors, which was given 1 h prior to virus adsorption and kept throughout the whole experiment. At 24 hpi, viral plaques were microscopically counted, and the number of plaques was plotted as a function of inhibitor concentration. Values are expressed as means ± SEM (error bars) of three independent experiments. Values are expressed as mean ± SEM of three independent experiments, * P < 0.05, ** P < 0.01, *** P < 0.001; one-way ANOVA followed by Bonferroni’s post test. ( C ) Representative images of infected HFFs (24 hpi) at an MOI of 1 PFU/cell and treated with HF4 (5 μM), CAY10727 (1 μM), or vehicle (DMSO). ( D ) Protein lysates from uninfected (mock) or infected HFFs (24 hpi) at an MOI of 1 PFU/cell treated with AFM30a (20 μM), HF4 (5 μM), GSK199 (20 μM) or vehicle (DMSO) were analyzed by immunoblotting to assess for viral expression with an anti-gD antibody; β-actin cellular expression was used for protein loading control. SH-SY5Y, ARPE-19, and HEK293 cells were infected with HSV-1 (MOI 1 PFU/cell) and then treated with increasing concentrations of HF4 (E) or CAY10727 (F) , which were given 1 h prior to virus adsorption and kept throughout the whole experiment. At 24 hpi, viral plaques were microscopically counted, and the number of plaques was plotted as a function of inhibitor concentration. Values are expressed as means ± SEM (error bars) of four independent experiments, * P < 0.05, ** P < 0.01, *** P < 0.001; one-way ANOVA followed by Bonferroni’s post test. (G) HFFs were silenced for PAD3 using specific siRNAs (siPAD3), as negative control cells were also similarly transfected with scrambled siRNA (siCTRL). At 24 h post-treatment (hpt), cells were infected with HSV-1 at an MOI of 1 PFU/cell. The efficiency of PAD3 protein depletion at 24 hpi was assessed by immunoblotting using antibodies against PAD3 or β-actin for equal loading. An anti-gD antibody was used to verify HSV-1 infection. Representative blots of three independent experiments are shown. ( H ) PAD3-silenced cells were infected with HSV-1 at an MOI of 1 PFU/cell. Viral supernatants were collected at 24 hpi and analyzed by standard plaque assay. Values are expressed as means ± SEM. Values are expressed as mean ± SEM of three independent experiments, *** P < 0.001; one-way ANOVA followed by Bonferroni’s post test. ( I ) Representative images of infected HFFs (24 hpi) at an MOI of 1 PFU/cell and transfected with the same siCTRL and siPAD3 described in the legend to .

Techniques Used: Infection, Virus, Adsorption, Concentration Assay, Western Blot, Expressing, Negative Control, Transfection, Plaque Assay

Synthesized:

Article Title: Reciprocal regulation of Th2 and Th17 cells by PAD2-mediated citrullination
Article Snippet: TGFβ1 (catalog 240-B-010), IL-6 (catalog 406-ML-005), and IL-23 (catalog 1887-ML-010/CF) were purchased from R&D Systems. .. Cl-am (synthesized in house), AFM30 (synthesized in house), and GSK199 (catalog 1549811-53-1, Cayman Chemical) were added at day 0 and maintained during the differentiation of Th cells. ..

Control:

Article Title: Regulation of Pseudomonas aeruginosa -Mediated Neutrophil Extracellular Traps
Article Snippet: .. Neutrophils were pre-treated with 10 μM diphenyleneiodonium chloride (DPI, Sigma Aldrich), pyrocatechol (100 μM, Sigma Aldrich), PAD inhibitors Cl-amidine (200 μM, Cayman Chemicals), GSK199 (10 μM, Cayman Chemicals), and GSK484 (10 μM, Cayman Chemicals) or DMSO control (0.01%) for 30 min at 37°C prior to treatment with PMA (100 nM) or different P. aeruginosa strains (MOI 10). ..

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(A) Supernatant HIV-1 RNA with PMAi and GSK484 treatments from entire cohort (n=30, study participant characteristics in Table S1). (B) Cell associated HIV-1 RNA in cohort stratified into in viremic and long-term ART suppressed groups. (C) Intact HIV-1 proviral reservoir from the two groups measured by IPDA. (D-E) HIV-1 5’ sLTR RNA (D) and Tat-Rev (E) quantified in viremic (n=14) and long-term ART suppressed groups (n=17). (F) PADI4 expression in the two groups normalised to RPP30. All statistical tests shown are Wilcoxon matched-pairs signed rank test * p<0.05, ** p<0.01, *** p<0.001.

Journal: bioRxiv

Article Title: De novo PADI4-mediated citrullination of histone H3 stimulates HIV-1 transcription

doi: 10.1101/2024.03.17.583304

Figure Lengend Snippet: (A) Supernatant HIV-1 RNA with PMAi and GSK484 treatments from entire cohort (n=30, study participant characteristics in Table S1). (B) Cell associated HIV-1 RNA in cohort stratified into in viremic and long-term ART suppressed groups. (C) Intact HIV-1 proviral reservoir from the two groups measured by IPDA. (D-E) HIV-1 5’ sLTR RNA (D) and Tat-Rev (E) quantified in viremic (n=14) and long-term ART suppressed groups (n=17). (F) PADI4 expression in the two groups normalised to RPP30. All statistical tests shown are Wilcoxon matched-pairs signed rank test * p<0.05, ** p<0.01, *** p<0.001.

Article Snippet: Padi4 Inhibitor GSK199 (MCE HY-103058) was added at a final concentration of 10 μM

Techniques: Expressing

(A)J-Lat 5A8 treated with PADI inhibitors and PMAi activation. GFP is measured by flow cytometry (n=8-11). (B)PADI4 expression in 5A8 measured by ddPCR under PMAi and GSK484 treatment (n=4). (C)Timecourse of 5A8 under PMAi and GSK484 treatment and flow cytometry quantification of GFP. Cells were washed and resuspended in new media without drugs after 1hr (n=3). (D)H3cit (R2, R8,R17 residues) quantification by immunoblot in 5A8 with PMAi and GSK484 treatment at two early timepoints (n=3). (E)T cell activation phenotype for 5A8. Cells gain CD69 during early T cell activation then CD25 during late T cell activation as shown by flow cytometry. (F)T cell activation status of 5A8 after 24 h with different PADI4 inhibitors and PMAi activated cells (n=3). (G)proliferation marker KI-67 staining of 5A8 treated for 24 h with PMAi and GSK484 (H)Polyclonal K562-Lat treated with PMAi and GSK484 for 24 h and quantified with flow cytometry (n=4). Data is shown as mean ± SEM. Statistical tests shown are unpaired T tests * p<0.05, ** p<0.01, *** p<0.001.

Journal: bioRxiv

Article Title: De novo PADI4-mediated citrullination of histone H3 stimulates HIV-1 transcription

doi: 10.1101/2024.03.17.583304

Figure Lengend Snippet: (A)J-Lat 5A8 treated with PADI inhibitors and PMAi activation. GFP is measured by flow cytometry (n=8-11). (B)PADI4 expression in 5A8 measured by ddPCR under PMAi and GSK484 treatment (n=4). (C)Timecourse of 5A8 under PMAi and GSK484 treatment and flow cytometry quantification of GFP. Cells were washed and resuspended in new media without drugs after 1hr (n=3). (D)H3cit (R2, R8,R17 residues) quantification by immunoblot in 5A8 with PMAi and GSK484 treatment at two early timepoints (n=3). (E)T cell activation phenotype for 5A8. Cells gain CD69 during early T cell activation then CD25 during late T cell activation as shown by flow cytometry. (F)T cell activation status of 5A8 after 24 h with different PADI4 inhibitors and PMAi activated cells (n=3). (G)proliferation marker KI-67 staining of 5A8 treated for 24 h with PMAi and GSK484 (H)Polyclonal K562-Lat treated with PMAi and GSK484 for 24 h and quantified with flow cytometry (n=4). Data is shown as mean ± SEM. Statistical tests shown are unpaired T tests * p<0.05, ** p<0.01, *** p<0.001.

Article Snippet: Padi4 Inhibitor GSK199 (MCE HY-103058) was added at a final concentration of 10 μM

Techniques: Activation Assay, Flow Cytometry, Expressing, Western Blot, Marker, Staining

Journal: PLOS Pathogens

Article Title: Citrullination profile analysis reveals peptidylarginine deaminase 3 as an HSV-1 target to dampen the activity of candidate antiviral restriction factors

doi: 10.1371/journal.ppat.1011849

Figure Lengend Snippet: HFFs were infected with HSV-1 (MOI 1) and then treated with increasing concentrations of HF4 (A) or CAY10727 (B) , two PAD3-specific inhibitors, which was given 1 h prior to virus adsorption and kept throughout the whole experiment. At 24 hpi, viral plaques were microscopically counted, and the number of plaques was plotted as a function of inhibitor concentration. Values are expressed as means ± SEM (error bars) of three independent experiments. Values are expressed as mean ± SEM of three independent experiments, * P < 0.05, ** P < 0.01, *** P < 0.001; one-way ANOVA followed by Bonferroni’s post test. ( C ) Representative images of infected HFFs (24 hpi) at an MOI of 1 PFU/cell and treated with HF4 (5 μM), CAY10727 (1 μM), or vehicle (DMSO). ( D ) Protein lysates from uninfected (mock) or infected HFFs (24 hpi) at an MOI of 1 PFU/cell treated with AFM30a (20 μM), HF4 (5 μM), GSK199 (20 μM) or vehicle (DMSO) were analyzed by immunoblotting to assess for viral expression with an anti-gD antibody; β-actin cellular expression was used for protein loading control. SH-SY5Y, ARPE-19, and HEK293 cells were infected with HSV-1 (MOI 1 PFU/cell) and then treated with increasing concentrations of HF4 (E) or CAY10727 (F) , which were given 1 h prior to virus adsorption and kept throughout the whole experiment. At 24 hpi, viral plaques were microscopically counted, and the number of plaques was plotted as a function of inhibitor concentration. Values are expressed as means ± SEM (error bars) of four independent experiments, * P < 0.05, ** P < 0.01, *** P < 0.001; one-way ANOVA followed by Bonferroni’s post test. (G) HFFs were silenced for PAD3 using specific siRNAs (siPAD3), as negative control cells were also similarly transfected with scrambled siRNA (siCTRL). At 24 h post-treatment (hpt), cells were infected with HSV-1 at an MOI of 1 PFU/cell. The efficiency of PAD3 protein depletion at 24 hpi was assessed by immunoblotting using antibodies against PAD3 or β-actin for equal loading. An anti-gD antibody was used to verify HSV-1 infection. Representative blots of three independent experiments are shown. ( H ) PAD3-silenced cells were infected with HSV-1 at an MOI of 1 PFU/cell. Viral supernatants were collected at 24 hpi and analyzed by standard plaque assay. Values are expressed as means ± SEM. Values are expressed as mean ± SEM of three independent experiments, *** P < 0.001; one-way ANOVA followed by Bonferroni’s post test. ( I ) Representative images of infected HFFs (24 hpi) at an MOI of 1 PFU/cell and transfected with the same siCTRL and siPAD3 described in the legend to .

Article Snippet: The PAD inhibitors Cl-amidine (Cl-A), BB-Cl-amidine (BB-Cl), GSK199, CAY10727, and AFM30a—also known as CAY10723—were obtained from Cayman Chemical (Ann Arbor).

Techniques: Infection, Virus, Adsorption, Concentration Assay, Western Blot, Expressing, Negative Control, Transfection, Plaque Assay

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gsk199 (Cayman Chemical)

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1) Product Images from "Citrullination profile analysis reveals peptidylarginine deaminase 3 as an HSV-1 target to dampen the activity of candidate antiviral restriction factors"

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