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. 2014 Nov 17;5:5375. doi: 10.1038/ncomms6375

Figure 3. Pathological changes in the lungs of BALB/c mice injected with omega-1, ipse or kappa-5 knockdown (kd) eggs of S. mansoni.

Figure 3

Test mice were injected into the lateral tail vein with 1,000 eggs previously transduced with lentiviruses containing gene-specific shRNAmir stems (omega-1, n=5; ipse, n=6; kappa-5; n=7). Control groups of mice received PBS (naive; n=6), 1,000 untreated wild type eggs (WT; n=7) or 1,000 eggs previously transduced with a lentivirus lacking the shRNAmir stem (that is, empty-vector (EV); n=6). All mice were euthanized 15 days after injection. (a) Total leukocyte numbers in cell suspensions from the right lung; cell populations: eosinophil granulocytes (EG), neutrophil granulocytes (NG), dendritic cells (DC), T-helper cells (Th), B cells (BC), interstitial macrophages (IM) or alveolar macrophages (AM). The percentage of leukocyte populations was established by flow cytometry (see Supplementary Fig. 2 for cell gating) and calculated as the total cell number of live cells isolated from the right lung from individual mice. Box plot representing the median and upper/lower quartile. Whiskers indicate the highest or lowest value. Kruskal–Wallis and Dunns post-hoc tests were used to establish statistical significance compared with the EV control: **P<0.01. (b) Effect of omega-1, ipse or kappa-5 knockdown of S. mansoni eggs on total serum IgE levels in mice 15 days following injection of eggs. (c) Effect of egg knockdown (kd) on granuloma formation in the left lung. Granuloma sizes were determined as the ratio of the granuloma area to the egg area. Scatter plot with mean±s.e.m. of the data pool representing all granulomata from all lungs from each experimental group (WT: n=48; EV: n=82; omega-1 kd: n=77; ipse kd: n=74; kappa-5 kd: n=84). Kruskal–Wallis and Dunns post-hoc tests were used to establish statistical significance compared with the EV control: *P<0.05, **P<0.01, ***P<0.001; not significant (ns). Numbers of mice: n=4 (WT); n=5 (EV, omega-1 kd); n=6 (ipse kd) and n=7 (kappa-5 kd). (d) Mean granuloma size in left lung sections from mice examined microscopically (original magnification: × 200), following staining with Masson–Goldner Trichrome. Scale bars, 100 μM. Arrows indicate S. mansoni eggs.