fitc conjugated integrin β3  (Bio-Rad)

Journal: bioRxiv

Article Title: A role for SHARPIN in platelet linear protein ubiquitination and function

doi: 10.1101/2021.01.13.426403

Figure Lengend Snippet: A – D) Washed wild-type and cpdm/cpdm platelets, +/- indomethacin (10 µM) and ARC66096 (1 µM), were stimulated for 10 min with thrombin (n = 11 mean ± s.e.) or CRP-XL (n = 11 mean ± s.e.) in the presence of PE-conjugated JON/A antibody directed against the high affinity form of integrin αIIbβ3 (A, C) and FITC-conjugated (Wug.E9) antibody (B, D) against the α-granule marker CD62P (P-selectin) before fixation and analysis. E & F) ATP release from washed wild-type and cpdm/cpdm platelets was monitored for 5 minutes in response to indicated concentrations of thrombin (n = 6 mean ± s.e.) or CRP-XL (n = 4 mean ± s.e.). Data expressed as % of ATP standard. Results were analysed by 2-way ANOVA with Bonferroni’s post-test. P values reported for genotype variable. α = 0.05. Star notation indicates individual post-test comparisons by genotype. **: p<0.01, ***: p<0.001.

Article Snippet: FITC-conjugated integrin β1 (CD29, HM beta 1-1, #MCA2298) and FITC-conjugated integrin β3 (CD61, #MCA2299) were from Bio-Rad (Oxford, UK).

Techniques: Marker

Journal: bioRxiv

Article Title: A role for SHARPIN in platelet linear protein ubiquitination and function

doi: 10.1101/2021.01.13.426403

Figure Lengend Snippet: Washed platelets were stimulated with the indicated agonists for 10 min in the presence of of PE-conjugated JON/A antibody directed against the high affinity form of integrin αIIbβ3 (A, C) and FITC-conjugated (Wug.E9) antibody (B, D) against the α-granule marker CD62P (P-selectin) before fixation and analysis. A & B) Integrin activation and α-granule secretion induced by ADP (10 µM) plus U46619 (30 µM) (n=7 mean ± s.e.). C) ATP release from washed wild-type and cpdm/cpdm platelets was monitored for 5 minutes in response to ADP (10µM)+U46619 (30µM) (n = 6 mean ± s.e.). Results in A - C were analysed by 2-way ANOVA with Bonferroni’s post-test. P values reported for genotype variable. α = 0.05. Star notation indicates individual post-test comparisons by genotype. Results in C were analysed by Wilcoxon test *: p<0.05, **: p<0.01, ***: p<0.001.

Article Snippet: FITC-conjugated integrin β1 (CD29, HM beta 1-1, #MCA2298) and FITC-conjugated integrin β3 (CD61, #MCA2299) were from Bio-Rad (Oxford, UK).

Techniques: Marker, Activation Assay

Journal: Scientific Reports

Article Title: TRAF3 negatively regulates platelet activation and thrombosis

doi: 10.1038/s41598-017-17189-1

Figure Lengend Snippet: Platelet secretion and aggregation are defective in the TRAF3 knockout mice. ( a ) Washed platelets from TRAF3 knockout mice (TRAF3 −/− ) or wild-type littermates (TRAF3 +/+ ) were stimulated with various concentrations of thrombin or collagen and simultaneously recorded for ATP secretion and aggregation. The aggregation and ATP release traces are representatives of at least three different experiments. ( b ) Aggregation and ATP secretion results in the experiments described in A were quantitated. ( c ) Total serotonin in 1 × 10 9 TRAF3 +/+ and TRAF3 −/− platelets from four mice with each genotype was measured by O-phthalaldehyde assay as described under “Experimental Procedures” (n = 4). ( d ) Detection of P-selectin in platelet lysates from TRAF3 +/+ and TRAF3 −/− mice by Western blot (n = 4).

Article Snippet: Fluorescein isothiocyanate (FITC)-conjugated rat anti-mouse P-selectin and integrin β3 antibodies were from BD Pharmingen.

Techniques: Knock-Out, Western Blot