Fraction of tumor kill, , vs. ratio of γδ T-cells to cancer cells, .

<p>Experiments <i>in vitro</i> (blue circles with SD; n = 3) and <i>in vivo</i> (red diamonds:  = 15, 30; red circles: ; red squares: ; with SD; n = 3 or 4; some error bars not visible at the scale of the figure). Mathematical model Eq. (1) with  = 3 µm and  = 7 µm <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0061398#pone.0061398-Jemal1" target="_blank">[1]</a>, <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0061398#pone.0061398-Arya1" target="_blank">[45]</a> (blue solid curve: least-squares fit to the in-vitro data; , <i>L</i> = 36 µm; <i>R</i>² = 0.99 and <i>p</i>-value = 0.0002 for only; red solid curve: least-square fit to the in-vivo data; , <i>L</i> = 679 µm; <i>R</i>² = 0.84 and <i>p</i>-value = 0.0037 for only; dashed blue curve: fit ignoring the outlier at  = 30; and ; dashed red curve: fit ignoring the outlier at  = 30; and <i>L</i> = 20µm). The fittings of the model, excluding the two outliers, are highly accurate in comparison with experimental data; in particular, the predicted diffusion penetration distance is much smaller <i>in vivo</i>.</p

Schematic of interaction of γδ T cells and cancer cells through IFN-γ.

<p>Schematic of interaction of γδ T cells and cancer cells through IFN-γ.</p

Automated quantitative determination of the amount of apoptosis in histological images.

<p><b>A.</b> A sample high power field image stained with apoptosis marker, CC3, shown in brown hues. <b>B.</b> Segmentation of CC3 positive regions (outlined in green) in the image in <b>A</b>.</p

Apoptosis calculation results.

<p>Apoptosis calculation results.</p

Expression of signaling molecules.

<p>(A) Western blot of angiogenesis related and connexin 43 proteins from each animal group. (B) Western blot of various molecules of oncogenic potential and relevant signaling pathways from each animal group.</p

Detection of stem cell homing.

<p>(A) Immunohistochemical detection was performed 36 hours after half a million of GFP overexpressed nanofiber expanded stem cells injected into the left ventricle of the rats, with fixed and paraffin embedded tissue sections from various organs using anti-GFP Ab (right panels) against respective controls (left panels) confirming stem cell homing to lung, spleen, liver and heart. (B) Heart sections were stained for human specific von Willebrand factor (vWF) and immunoglobulin G (IgG) as a control.</p

Expression of gap junctional protein, connexin 43.

<p>Heart tissue sections were stained for gap junctional protein, connexin 43 from all four groups using immunohistochemistry. Red color indicates connexin 43 stain and blue color indicates DAPI, a nuclear stain. Merged connexin 43 and DAPI of each group is displayed in micrographs.</p

Evaluation of cardiac function by echocardiography.

<p>(A) Representative M-mode echocardiographic images after six weeks of therapy demonstrating improved anterior wall motion in the VIP > Exp > FCB > Med groups. Inset shows the short axis two-dimensional image of the LV. (B) Echocardiographic parameters of cardiac function using fractional shortening (FSa), anterior wall tissue velocity (TVa) and wall motion score index (WMSI) plotted as a function of time within each group demonstrating significant improvement in function in the VIP group compared to controls. (C) Strain and strain rates presented as a function of time within each group with significant improvement in the VIP group.</p

Evaluation of neovascularization.

<p>Cyro-preserved sections were stained with alkaline phosphatase, and were evaluated capillary density area (%) per high power field (A), and total capillary number per high power field (B) in histochemistry images. Representative images of each group (C) were shown in the lower panel. Capillary count and total capillary areas (A & B) were measured by Image Quant software demonstrating highest capillary area density in the VIP group.</p

Evaluation of total ischemic area using Masson's trichrome staining.

<p>Paraffin-blocked tissues from infarct zones were sectioned and stained with Masson's trichrome after six weeks of therapy demonstrating decreased infarct size in VIP < Exp < FCB < Med groups. Five sections were examined from each heart. Four hearts from each group were evaluated. Every section was examined with low power field to visualize gross heart morphology, and to visualize fibrous tissues after ischemia, and area of interest of each heart section is magnified. Representative gross heart pictures are shown here with areas of interest in higher magnification.</p