The cell culturemediumwas DMEM for growing SK Deborah BE2 cell line and was RPMI 1640 for growing SH SY5Y cell line. Cellswere growntill 80%confluency, preceding todrug treatment and then starved within their respective cell culturemediumcontaining 2% FBS for 24 h. The Bcl 2 inhibitor CTEP GluR Chemical and genistein were acquired. Drugs were dissolved in dimethyl sulfoxide tomake a stock option and aliquots were stored at 20 C until ready for use. Doseresponse studies were performed to determine the suitable doses of the drugs for induction of apoptotic death. Cell viability was determined utilizing an MTT colorimetric assay system. The essential principle of this analysis would be to measure the action of mitochondrial enzyme system that turns yellow MTT to purple colored formazan. SH SY5Y cells and both SK Deborah BE2 were seeded at 3?105 cells/well in two 96 well plates separately. Different doses of GST and HA and their combination were added to each plate in triplicates and plates were incubated overnight in a humidified incubator containing five minutes CO2 at 37 C. Then, MTT reagent was added in each plate and incubated for 4 h at 3-7 C. Formazan precipitate was dissolved by pipetting each well up and down with 100 ul of isopropyl alcohol. Plates were read on the DU800 spectrophotometer using 570 nm since the test wavelength. Cell viability data were analyzed using CompuSyn pc software to ascertain a combination index for synergism in drug combination studies. Traditionally, CI 1 indicates antagonism, CI_1 Plastid indicates additive effect, and CI 1 indicates synergism in the effective doses. We observed a low CI using 10 uM HA 250 uM GST in SK Deborah BE2 cell line and also a low CI using 5 uM HA 100 uM GST in SH SY5Y cell line. Therefore, these particular doses of the medications and their combinations were selected due to their complete inhibitory activity on cell growth in all other studies. Both cell lines in culture dishes were handled with HA, GST, and HA GST for 2-4 h and examined under the phase contrast microscope. Canagliflozin clinical trial Treatments caused different morphological features of apoptosis in cells on the plates. Using phase contrast microscopy, black and white photographs were taken. Further, cells from each treatment were washed with PBS and sedimented on slides by using the Eppendorf 5804R centrifuge at 106 g for 5 min. Cells were stained with Wright discoloration and set with 95% ethanol. Morphological features of apoptotic cells were discovered under the light microscope. Morphological characteristics of apoptosis involved reduced amount of chromatin condensation, cell volume, and existence of membranebound apoptotic bodies. Four randomly selected areas were measured for at least 800 cells. The proportion of apoptotic cells was calculated from three separate studies.