Fig. 2

Blast cell differentiation potential of esculetin in Kasumi-1 cells. A Microscopic analysis of NBT stained cells. Exponentially growing Kasumi-1 cells treated with/without esculetin (100 µM) for 96 h. PMA was used for ROS stimulation and MTT was added. Manual counting (Lower left panel) and colorimetric analysis of dissolved crystals in DMSO (lower right panel) of treated/untreated cells was carried out to determine relative staining using NBT reduction assay (**p ≤ 0.02). B Flow cytometric analysis of unfixed/unstained Kasumi-1 cells treated with varied esculetin concentrations to measure cellular granularity (FSC versus SSC) (Upper panel) and cell size (FSC verses cell count) (Lower Panel). High side scatter (SSC) indicative of cells with enhanced granularity. A dose dependent left shift of the forwarded scatter (FSC) upon esculetin treatment indicative of reduction in cell size. Scatter characteristics of 10,000 cells were analysed. Percentage of shifted cells was measured using flowing software. C Surface expression analysis of immaturity markers (CD34) (Upper Panel) and maturity marker (CD38) (Lower panel) using flow cytometry following 96 h of esculetin treatment. Percentage of shifted cells was measured using flowing software