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Figure 3. Comparative study of expression of c-kit in HSCs of five experimental animal groups, viz., the 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 c-kit 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 c-kit 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 c-kit percent positive cells in the high density compartments (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). Comparisonof Distribution of c-kit⁺ Cells between LDC and HDC. The line diagram shows both the LDC and HDC HSCs there is a steep high rise in their expression level of c-kit cells after T11TS administration, but compared to HDC the LDC cells shows the high level of proliferation as compared to ENU and normal since c-kit ligation opens up SCF signaling cascade which leads to lineage commitment of HSCs, so the results obtained here with also hints driving of both the group of cells towards lineage commitment; C: representative images (100× magnification) showing immunofluorescent staining of cell surface c-kit 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) in both the LDC and HDC compartment. Each condition was observed in triplicate and six images were taken for each sample. Figures are representative of the group. Results were expressed as the percent c-kit-positive cells out of the total number of cells counted for each experimental group. Column C-kit: FITC-stained c-kit expression in bone marrow derived hematopoietic stem cells green in color due to the presence of c-kit. Column Dapi: Nuclei appear blue due to staining with DAPI. Column Merge: Merged image FITC-stained 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. At the bottom the comparison of the mean intensity of c-kit positive cells between LDC and HDC has been also depicted. *Significant (P < 0.001) decrease in ENU compared with that of normal control group. #Significant (P < 0.001) increase, when individually comparing T11TS treated groups with glioma-bearing ENU group in both the compartments. HSCs: hematopoietic stem cells; LDC: low density compartment; HDC: high density compartments; ENU: N-ethyl-N-nitrosourea