puromycin. Pool puromycin resistant HT-29 cells were then treated with the study drug, and analyzed Dihydromyricetin by confocal microscopy. The confocal fluorescence microscopy for GFP transduced with lentiviral LC3B LC3B GFP construct were fixed with paraformaldehyde third Fluorescence signals were visualized and recorded through a microscope LSM 5 Pascal laser scanning used with appropriate filter combinations and detection according to the spectrum of the fluorochrome. Acridine orange to demonstrate autophagy After drug treatment, acridine orange was the culture medium and the cells were incubated at 37 for 15 min 30th The cells were then trypsinized and. With cold PBS 2 and under a confocal microscope Fluorescence with a bandpass filter 490 nm, and the blue fluorescence of the red and green channel recorded were excited and merge. A change from green to red fluorescence vesicles S acids With autolysosomes. In the presence of bafilomycin A1, an inhibitor that Bl Cke lysosome fusion with lysosomes autophagosome fluorescence was observed not only green red, and this treatment was used as a negative control for the F Used coloring. Protein samples Western blots were in a lysis buffer, prepared with standardized measurement NanoDrop and boiled in LDS sample buffer. The samples were then loaded onto SDS-PAGE gels with 14 electrophoretic transfer onto a polyvinylidene fluoride membrane. Western blotting was performed as previously described, 44, and F Staining was performed using the software Image J.
All experiments were repeated at least twice and the mean values and standard deviations were calculated from triplicate experiments. Mitochondrial release of cytochrome c mitochondrial After drug treatment, the release of cytochrome c were analyzed using a selective digitonin lysis method as previously described.44 an antique Body against subunit IV of cytochrome oxidase was used as marker of the cytosolic fraction. Calculating the combination index interaction between celecoxib and ABT 737 once with CalcuSyn as previously reported.44 Statistical Resveratrol analysis The statistical significance of differences between the experimental variables and the reference group was determined using Pr Examination results r. P 0.05 was considered statistically significant. Values repr Sentieren the mean SD of three experiments. A. INTRODUCTION herk mmlichen Not stero Ans inflammatory stero Ans such as aspirin, naproxen and ibuprofen is known that second the inflammation by blocking the formation of prostaglandins and thromboxanes through inhibition of cyclooxygenase-1 and COX-Break To block their effectiveness or effects on the heart-piece and their F Ability, the action of COX-1 and COX-2, or both. Anti-inflammatory and increased ht The therapeutic value of these inhibitors is believed to result primarily from the inhibition of COX-2, w Occur while the side effects of bleeding and stomach ulcers due to inhibition of COX first An increase increase Expression of COX-2 in inflamed tissue by an increase Increase its decay product, PGE2, the peripheral nociceptors terminals pain accompanied sensitized. Highly selective COX-2 inhibitors such as coxibs have anti-cancer and anti-inflammatory activity How it is Under