Fig. 1
Deletion of mTOR protein in platelets. a Genotyping of mTORfl/fl, mTORfl/wt PF4-Cre + (mTOR+/−) and mTORfl/fl PF4-Cre + (mTOR−/−) mice using polymerase chain reaction. The top and bottom images were cropped from different gels and full-length/original gels were shown in Additional file 3: Part III. b Lysates from PF4-Cre− (wildtype, WT, W) littermate, or mTOR−/− (KO, K) platelets were immunoblotted with mTOR, Raptor, Rictor, and Actin antibodies, these images (separated by horizontal white space) were cropped from the different/same gels and full-length/original blots were shown in Additional file 3: Part III. c Flow cytometric analysis of the surface expression of CD41 (αIIbβ3), CD42b (GPIbα), and GPVI of resting WT and mTOR−/− murine platelets. Results are expressed as mean fluoresce intensity (MFI) ± SEM (n = 3). d–f Representative photomicrographs and quantification of the adhesion and aggregation of WT or mTOR−/− platelets on 20 μg/mL or g–i 50 μg/mL type I fibrillar collagen-coated surface. The fluorescent platelets adhering to coated Bioflux micro flow chambers were recorded by video (exposure time were fixed as 1.5 or 0.8 s, respectively). The coverage surface area and total integrated fluorescence were quantified by Bioflux software (Fluxion) as a measure of platelet thrombi formation. Quantified results are expressed as the mean percentage of surface coverage (left panels) or mean integrated fluorescence intensity (right panels) ± SEM (n ≥ 3 per group, * indicates P < 0.05, paired Student’s t test)