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Figure 4. Platelets express the bile acid receptor TGR5. Human platelets lysates (40×106) were analyzed regarding their protein expression of TGR5 (A) with and without CRP stimulation. Lysates of human <t>gallbladder</t> tissue and TGR5 transfected HEK293 cells served as positive control. (B) Quantification of TGR5 expression in human platelets lysates (n = 3). (C) Human platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and Actin filaments (n = 3). (D) Analysis of cDNA isolated from murine platelets were analyzed regarding the expression of TGR5 mRNA (n = 3). (E) Murine platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and actin filaments (n = 3). (F) Murine platelets of wildtype and TGR5-/- mice were pre-incubated with TCDC, TLC or DMSO control for 1 h. After preincubation, platelets were allowed to adhere on a fibrinogen matrix for 60 min and total adhesion was recorded (n = 3–6). Surface coverage and number of adherent cells (E) was determined using Fiji software. Statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test. *p < .05, *p < .001. Data shown as mean + SEM.
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Figure 4. Platelets express the bile acid receptor TGR5. Human platelets lysates (40×106) were analyzed regarding their protein expression of TGR5 (A) with and without CRP stimulation. Lysates of human <t>gallbladder</t> tissue and TGR5 transfected HEK293 cells served as positive control. (B) Quantification of TGR5 expression in human platelets lysates (n = 3). (C) Human platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and Actin filaments (n = 3). (D) Analysis of cDNA isolated from murine platelets were analyzed regarding the expression of TGR5 mRNA (n = 3). (E) Murine platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and actin filaments (n = 3). (F) Murine platelets of wildtype and TGR5-/- mice were pre-incubated with TCDC, TLC or DMSO control for 1 h. After preincubation, platelets were allowed to adhere on a fibrinogen matrix for 60 min and total adhesion was recorded (n = 3–6). Surface coverage and number of adherent cells (E) was determined using Fiji software. Statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test. *p < .05, *p < .001. Data shown as mean + SEM.
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Figure 4. Platelets express the bile acid receptor TGR5. Human platelets lysates (40×106) were analyzed regarding their protein expression of TGR5 (A) with and without CRP stimulation. Lysates of human <t>gallbladder</t> tissue and TGR5 transfected HEK293 cells served as positive control. (B) Quantification of TGR5 expression in human platelets lysates (n = 3). (C) Human platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and Actin filaments (n = 3). (D) Analysis of cDNA isolated from murine platelets were analyzed regarding the expression of TGR5 mRNA (n = 3). (E) Murine platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and actin filaments (n = 3). (F) Murine platelets of wildtype and TGR5-/- mice were pre-incubated with TCDC, TLC or DMSO control for 1 h. After preincubation, platelets were allowed to adhere on a fibrinogen matrix for 60 min and total adhesion was recorded (n = 3–6). Surface coverage and number of adherent cells (E) was determined using Fiji software. Statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test. *p < .05, *p < .001. Data shown as mean + SEM.
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Figure 4. Platelets express the bile acid receptor TGR5. Human platelets lysates (40×106) were analyzed regarding their protein expression of TGR5 (A) with and without CRP stimulation. Lysates of human <t>gallbladder</t> tissue and TGR5 transfected HEK293 cells served as positive control. (B) Quantification of TGR5 expression in human platelets lysates (n = 3). (C) Human platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and Actin filaments (n = 3). (D) Analysis of cDNA isolated from murine platelets were analyzed regarding the expression of TGR5 mRNA (n = 3). (E) Murine platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and actin filaments (n = 3). (F) Murine platelets of wildtype and TGR5-/- mice were pre-incubated with TCDC, TLC or DMSO control for 1 h. After preincubation, platelets were allowed to adhere on a fibrinogen matrix for 60 min and total adhesion was recorded (n = 3–6). Surface coverage and number of adherent cells (E) was determined using Fiji software. Statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test. *p < .05, *p < .001. Data shown as mean + SEM.
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Figure 4. Platelets express the bile acid receptor TGR5. Human platelets lysates (40×106) were analyzed regarding their protein expression of TGR5 (A) with and without CRP stimulation. Lysates of human <t>gallbladder</t> tissue and TGR5 transfected HEK293 cells served as positive control. (B) Quantification of TGR5 expression in human platelets lysates (n = 3). (C) Human platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and Actin filaments (n = 3). (D) Analysis of cDNA isolated from murine platelets were analyzed regarding the expression of TGR5 mRNA (n = 3). (E) Murine platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and actin filaments (n = 3). (F) Murine platelets of wildtype and TGR5-/- mice were pre-incubated with TCDC, TLC or DMSO control for 1 h. After preincubation, platelets were allowed to adhere on a fibrinogen matrix for 60 min and total adhesion was recorded (n = 3–6). Surface coverage and number of adherent cells (E) was determined using Fiji software. Statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test. *p < .05, *p < .001. Data shown as mean + SEM.
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Figure 4. Platelets express the bile acid receptor TGR5. Human platelets lysates (40×106) were analyzed regarding their protein expression of TGR5 (A) with and without CRP stimulation. Lysates of human <t>gallbladder</t> tissue and TGR5 transfected HEK293 cells served as positive control. (B) Quantification of TGR5 expression in human platelets lysates (n = 3). (C) Human platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and Actin filaments (n = 3). (D) Analysis of cDNA isolated from murine platelets were analyzed regarding the expression of TGR5 mRNA (n = 3). (E) Murine platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and actin filaments (n = 3). (F) Murine platelets of wildtype and TGR5-/- mice were pre-incubated with TCDC, TLC or DMSO control for 1 h. After preincubation, platelets were allowed to adhere on a fibrinogen matrix for 60 min and total adhesion was recorded (n = 3–6). Surface coverage and number of adherent cells (E) was determined using Fiji software. Statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test. *p < .05, *p < .001. Data shown as mean + SEM.
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Figure 4. Platelets express the bile acid receptor TGR5. Human platelets lysates (40×106) were analyzed regarding their protein expression of TGR5 (A) with and without CRP stimulation. Lysates of human gallbladder tissue and TGR5 transfected HEK293 cells served as positive control. (B) Quantification of TGR5 expression in human platelets lysates (n = 3). (C) Human platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and Actin filaments (n = 3). (D) Analysis of cDNA isolated from murine platelets were analyzed regarding the expression of TGR5 mRNA (n = 3). (E) Murine platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and actin filaments (n = 3). (F) Murine platelets of wildtype and TGR5-/- mice were pre-incubated with TCDC, TLC or DMSO control for 1 h. After preincubation, platelets were allowed to adhere on a fibrinogen matrix for 60 min and total adhesion was recorded (n = 3–6). Surface coverage and number of adherent cells (E) was determined using Fiji software. Statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test. *p < .05, *p < .001. Data shown as mean + SEM.

Journal: Platelets

Article Title: The bile acid receptor TGR5 inhibits platelet activation and thrombus formation.

doi: 10.1080/09537104.2024.2322733

Figure Lengend Snippet: Figure 4. Platelets express the bile acid receptor TGR5. Human platelets lysates (40×106) were analyzed regarding their protein expression of TGR5 (A) with and without CRP stimulation. Lysates of human gallbladder tissue and TGR5 transfected HEK293 cells served as positive control. (B) Quantification of TGR5 expression in human platelets lysates (n = 3). (C) Human platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and Actin filaments (n = 3). (D) Analysis of cDNA isolated from murine platelets were analyzed regarding the expression of TGR5 mRNA (n = 3). (E) Murine platelets were allowed to adhere on a fibrinogen matrix and were stained for TGR5 and actin filaments (n = 3). (F) Murine platelets of wildtype and TGR5-/- mice were pre-incubated with TCDC, TLC or DMSO control for 1 h. After preincubation, platelets were allowed to adhere on a fibrinogen matrix for 60 min and total adhesion was recorded (n = 3–6). Surface coverage and number of adherent cells (E) was determined using Fiji software. Statistical significance was determined by two-way ANOVA with Tukey’s multiple comparisons test. *p < .05, *p < .001. Data shown as mean + SEM.

Article Snippet: For immunoblotting targeting human TGR5, an antiTGR5 antiserum (M39) was raised against amino acids 298–318 of human TGR5 (NP_733800) and characterized using TGR5YFP transfected HEK293 cells and human gallbladder tissue as previously described.19 Glyceraldehyde 3-phosphate dehydrogenase (GAPDH, # Sc -32233, Santa Cruz Biotechnology, 1:10000) and horseradish peroxidase-conjugated anti-mouse secondary antibody (#P0447, DAKO, 1:5000) served as loading control.

Techniques: Expressing, Transfection, Positive Control, Staining, Isolation, Incubation, Control, Software