The role of tissue-mediated platelet clearance in L donovani–induced thrombocytopenia. Splenectomized and sham operated mice were infected with L donovani amastigotes for 28 days, similar to WT mice, and bled for the blood counts. (A) Platelet count and MPV of the study groups (n = 15 for each group). Data were pooled from 3 independent experiments and analyzed using ANOVA with the post hoc Tukey’s test comparing mean ± SD of all groups. (B) Platelet count and MPV of sham/infected (n = 4) and splenectomized/infected (n = 5) mice after administration of exogenous rTPO (50 µg/g bodyweight) for 5 consecutive days. Data are representative of a single experiment and analyzed with the unpaired Student t test comparing the mean ± SD between 2 groups. (C) Flow cytometric analysis was performed for the estimation of IgG- and IgM-bound platelets in the circulation of infected mice. Representative histograms show the gating strategy for IgG- and IgM-bound platelets in uninfected and infected mice. (D) Dot plots represent the percentage of total platelets with bound IgG and IgM in uninfected (n = 5) and infected (n = 15) mouse platelets. Data are representative of 2 independent experiments and were analyzed with the unpaired Student t test comparing the mean ± SD between the 2 groups. (E) Flow cytometric analysis was performed on the freshly isolated platelets for the estimation of desialylated platelets in circulation. Histograms representative of gating strategy for lectin⁺ platelets in infected (i) and uninfected control (c) mice. Neuraminidase-treated platelets (n) were used as the positive control for each lectin. (F) Percentage of RCA-1 and MAL-2 lectin⁺ platelets from uninfected (n = 4) and infected (n = 15) mice. Data were pooled from 2 independent experiments and analyzed using the unpaired Student t test comparing the mean ± SD between 2 groups. *P < .05; **P < .01; ***P < .001; ****P < .0001.