Fig. 1

L. amazonensis proliferation in BMFs in the presence or absence of osteopontin. L. amazonensis amastigotes at a ratio 4:1, were added or not (Ctrl) to (a) C57BL/6^+/+ or (b) opn mutant (C57BL/6^−/−), representative images of BMFs populations. Twenty four and 48 h later, each BMFs population was analysed by immunostaining. Nuclei were stained with Hoechst (blue), vacuoles with Lysosome-associated membrane protein Lamp-1 Ab and FITC-labelled conjugate (green) and amastigotes with 2A3-A26 Ab and Texas Red-conjugate (red). All images are in phase-contrast optical microscopy. c. Parasite Intensity: Mean intensities and intracellular parasite crowding of L. amazonensis amastigotes per infected cell was monitored by manual analysis of immunofluorescence image captures (AxioVision) of at least 2 different experiments (≥ 40 cells evaluated per condition). Statistical analyses were performed using the QP3.0 program designed for Quantitative parasitology as described in the Methods section. Mean intensities were compared by the Bootstrap test and 2-sided bootstrap p-values are given as follows: WT vs KO at 24 h, NS; WT vs KO 48 h, P = 0.0001; WT 24 h vs 48 h and KO 24 h vs 48 h, P = 0.00001. Mean crowding across samples was significant with non-overlapping confidence limits (Cl) at 97.5% and a p-value< 0.0595%, (95% Cl). Statistics are analytically presented on Additional file 2: Table S1A and B