Additional file 2: Figure S2. of Pasteurella multocida toxin- induced osteoclastogenesis requires mTOR activation

(a) PMT-mediated miR-21 activation is mTOR-independent. MiR-21 expression of PMT-stimulated cells was determined by RT-PCR and the effects of mTOR pathway inhibition by rapamycin (10 ng/ml) were addressed (normalisation to RnU6b). The indicated standard deviation was calculated from three independent experiments (mean ± SD; n = 3). Statistical analysis was performed using an unpaired Student’s t-test (***p ≤ 0.005). (b) p-STAT-3 (Tyr705) and STAT-3 was detected by immunoblotting as described above. (c) Uptake of the fluorescent 5’6-FAM antagomiR 21 was detected 24 h after transfection by fluorescent microscopy. As controls, untransfected cells and cells transfected with non = fluorescent control antogomiR were used (n = 3). (d) PDCD4 expression was monitored by western blotting in cells 48 h after transfection with a antagomiR 21 or control antagomiR in the presence or absence of a PMT stimulus (12 h at 5 nM). (JPEG 798 kb)