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Vehicle treated control 3. D ) in MSE and MIT treated SH-SY5Y cells as determined by the Trypan blue exclusion assay. Values are the mean of quadruplet cultures of MSE experiment and duplicate cultures of MIT experiment. Bars are SEM. ANOVA with Dunnet post test.

LDH released into the culture medium is measured with a 10-minute coupled enzymatic

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assay that results in the conversion of resazurin into resorufin. For cytotoxicity assay; MSE treated HepG2 cells were cultured as described in section 2. C for 10 min.

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MSE and MIT. From these estimates it appears that the SH-SY5Y cells are the most sensitive of those examined to the cytotoxic and possibly cytostatic effect of MSE. Based upon my estimation of 42% MIT-like compound in MSE extract the SHSY5Y cell IC50 for MSE is equal to 9. M MIT-like compound. This is captain kratom powder review not dissimilar to the experimentally determined IC50 for pure MIT of 7. To assess the long-term effect of MSE on surviving cells after acute treatment a clonogenicity assay was performed after 24 hr treatment on HEK 293 and SHSY5Y cells.

Annexin V conjugate assay for apoptosis detection 5. A possible role of caspases in MSE and MIT induced cell death 5. Possible involvement of pro-apoptotic caspases (8 and 9) 5. Possible involvement of caspases executor (3 and 7) 5. ROS generation in SH-SY5Y cells treated with MSE and MIT 5.

In the absence of FBS (Panel A) the SH-SY5Y cells failed to proliferate or migrate into the wound area (refer to fig. In the presence of FBS (Panel B) it can clearly be seen that the cells proliferated and migrated into the wound area. In the presence of MSE (without FBS) no proliferation or migration was observed (Panels CD E and F).

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