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Figure 2. Comparative study of expression of Sca-1 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 Sca-1 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 Sca-1 percent positive cells in the low density compartment 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 Sca-1 percent positive cells in the high density compartment (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 Sca-1⁺ cells between LDC and HDC: The line diagram shows both the LDC and HDC HSCs there are a steep high rise in their expression level of Sca-1 cells after T11TS administration, but compared to HDC the LDC cells show the high level of proliferation as compared to ENU and Normal groups probably hinting activated state of HSCs towards further maturity since Sca-1 expressed in both primitive and mature cells; C: Expression of Sca-1 in LDC was analyzed by immunoblotting using anti-Sca-1 specific antibody. The immunoblot shows band intensities for the Sca-1. β-actin was used as loading control and blots were reprobed with anti-β actin antibody to establish equivalent loading. Bands were individually analyzed densitometrically and relative pixel intensities of individual, group were displayed in bar diagrams; D: expression of Sca-1 in HDC was analyzed by immunoblotting using anti-Sca-1 specific antibody. The immunoblot shows band intensities for the Sca-1. β-actin was used as loading control and blots were reprobed with anti-β actin antibody to establish equivalent loading. Bands were individually analyzed densitometrically and relative pixel intensities of individual, group were displayed in bar diagrams. ^*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. At the bottom, the comparison of expression of relative pixel intensities of Sca-1⁺ cells between LDC and HDC. HSCs: hematopoietic stem cells; LDC: low density compartment; HDC: high density compartments; ENU: N-ethyl-N-nitrosourea