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Figure 5. Comparative study of expression of Tie-2 in HSCs of five experimental animal groups, viz., normal, healthy control rats (N), glioma-bearing rats (ENU), and rats having received the first (ET1), second (ET2) and third (ET3) doses of T11TS. Flowcytometric studies of the expression of Tie-2 in HSCs cells isolated from bone marrow of normal control and of glioma-bearing rats before and after T11TS treatment have been shown. A: Flowcytometric analysis of Tie-2 percent positive cells in the LDC using BD Cell Quest Pro Software by using scatter from a single representative experiment. The percentage of expression refers to the percent positive cells out of 10,000 cells analyzed represented in graphical form in bar diagram. Column values represent mean ± SD (n = 6 animals per group); B: Flowcytometric analysis of Tie-2 percent positive cells in the HDC using BD Cell Quest Pro Software by using scatter from a single representative experiment. The percentage of expression refers to the percent positive cells out of 10,000 cells analyzed represented in graphical form in bar diagram. Column values represent mean ± SD (n = 6 animals per group). Comparison of Distribution of Tie-2⁺ Cells between LDC and HDC: Tie-2 which is ligand to Ang-1 and is expressed on HSCs is highly expressed in the LDC group compared to the HDC in ENU treated groups, though both groups show up-regulated expression compared to the normal group. This shows that the immature HSCs are more protected than the mature cells at HDC during the ENU insult. T11Ts down regulates the higher expression of both the groups below normal levels, indicating that T11TS revive the quiescent state of HSCs driving them towards differentiation and self-renewal properties; C: representative images (100× magnification) showing immunofluorescent staining of Tie-2 expression on bone marrow derived hematopoietic stem cells of normal (N) and glioma-bearing rats before (ENU) and after T11TS treatment (ET1, ET2 and ET3 groups) in both the groups LDC and HDC. Each condition was observed in triplicate and six images were taken for each sample. Column Tie-2: TRITC stained Tie-2 expression in isolated BMHSC cells which appears red in color. Column Dapi: DAPI-stained nuclei appear blue. Column Merge: Merged picture TRITC fluorescence intensity of each group was analyzed with Nikon's Nis - Elements D3.00 software and the mean intensity was expressed in bar diagrams. Individual bar values represent mean intensity ± SD of the respective group. ^*Significant (P < 0.001) increase in ENU compared with that of normal control group. ^#Significant (P < 0.001) decrease, when individually comparing T11TS treated groups with glioma-bearing ENU group. At the bottom the comparison of the mean intensity of Tie-2 positive cells between LDC and HDC has been also depicted. HSCs: hematopoietic stem cells; LDC: low density compartment; HDC: high density compartments; ENU: N-ethyl-N-nitrosourea