Figure 2

Homing of Ad-MSCs to irradiated lung.

(A) Electrophesis in 1.5% agarose gel for verifying the specificity of hBeta2-MG primers to the human cells. (B) Expression of hBeta2-MG in lung tissue within 14 days post-irradiation. (C) Expression of rSDF-1α in lung tissue at 3 days post-irradiation. The expression levels of genes, including hBeta2-MG and rSDF-1α, were tested using quantitative real-time PCR. Rat Actin (rActin) was set as the internal control for determining ΔC_T values. Fold increases in expression were normalized to normal group by determining 2^−ΔΔCT values. Data shown are the mean ± S.D. of nine independent samples. This experiment was performed only once. One-way ANOVA analysis was used for comparing the differences of data among groups. *P ≤ 0.05 and **P ≤ 0.01 (significant high) versus the Normal + Ad-MSC group. ns: no statistical significance. (D) IHC-staining for Beta2-MG only in human cells and for SDF-1α in lung tissues at 3 days post-irradiation. Left rank: Magnification at 200×. Scale bar, 100 μm. Right rank: Magnification at 1000×. Scale bar, 20 μm. Black arrow: hBeta2-MG-positive cells. (E) IF-staining for CXCR4 in Ad-MSCs. Left: Propidium iodide for nuclear; Middle: Alexa Fluor 488 for detecting CXCR4 in Ad-MSCs; Right: Overlay. Magnification at 2000×. Scale bar, 10 μm. (F) FACS analysis for CD4⁺ or CD8⁺ T lymphocytes in peripheral blood samples at 14 days post-irradiation. (G) Percent of CD4⁺ or CD8⁺ T lymphocytes in peripheral blood monocytes (PBMCs). Upper: Percent of CD4⁺/CD8⁻ T lymphocytes; Lower: Percent of CD8⁺/CD4⁻ T lymphocytes. Data shown are the mean ± S.D. of nine independent samples. This experiment was performed only once. One-way ANOVA analysis was used for comparing the differences of data among groups. ^$P ≤ 0.05 and ^$$P ≤ 0.01 (significant low) versus the Normal group, 15 Gy group and Normal + Ad-MSC group.