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. 2021 Jun 7;12:693974. doi: 10.3389/fimmu.2021.693974

Figure 2.

Figure 2

Platelet CLEC-2 upregulates podoplanin and CD44 expression on LPS-stimulated BMDM. (A) BMDM were incubated in the presence or absence of LPS (1µg/ml) for 24h. (B, C) LPS-stimulated BMDM (Mϕ) were co-cultured in the absence or presence of WT or CLEC-2-deficient platelets (100 platelets: 1 Mϕ). (A–C) Median of fluorescence intensity (MFI) of podoplanin (n=4) and CD44 (n=3) was detected by flow cytometry. (D) Mϕ were cultured on glass, and WT or CLEC-2-deficient platelets were added for 1h. Platelets (CD41, green), podoplanin (red) and CD44 (cyan) were detected using confocal microscopy. Images are representative of 4 independent experiments. (E) Cell area and (F) circularity were analysed using ImageJ. (G) Mϕ were cultured on glass in the presence or absence of WT platelets (red) for 1h, before fixing and imaging by electron microscopy. The statistical significance between 2 groups was analyzed using a student’s paired t-test and the statistical difference between multiple groups using one-way ANOVA with Tukey’s multiple comparisons test. *p < 0.05, ***p < 0.001, ****p < 0.0001.