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. 2023 Apr 7;6:376. doi: 10.1038/s42003-023-04766-6

Fig. 4. Divalent AYP1 and podoplanin do not activate human platelets.

Fig. 4

Platelet aggregation was monitored by light transmission aggregometry at 37 °C with constant stirring at 1200 rpm for 5 mins. a Representative traces of human platelet aggregation (2 × 108 platelets/ml) induced by monoclonal antibody (mAb) AYP1 (66 nM) in the absence and presence of the anti-FcγRIIA IV.3 Fab (30 µg/ml), AYP1 F(ab’)2 (66 nM) and AYP1 Fab fragments (66 nM). Platelets were preincubated with IV.3 Fab for 10 min at 37 °C prior to stimulation. b Percentage of platelet aggregation induced by mAb AYP1 or AYP1 Fab fragments (n = 3 biologically independent experiments). c Representative traces of platelet aggregation following incubation with recombinant Fc-podoplanin (rFc-PDPN,136 nM), HEK293T PDPN−/− cells (5 × 105 cells) or wild-type HEK293T cells (5 × 105 cells). d Percentage of platelet aggregation induced by rFc-PDPN, HEK293T PDPN−/− cells or wild-type HEK293T cells (n = 3 biologically independent experiments). Significance was measured using one-way ANOVA with a Bonferroni post-hoc test where P ≤ 0.05. Data presented as mean ± SD.