AO/EB dual staining: Reduced programmed cell death signs by statistically optimized aqueous extracts of <i>T</i>.<i>terrestris</i> in oxalate injured NRK-52E renal epithelial cells.

<p>Detection of apoptosis, necrosis and quantitative changes in dual stained (AO/EB) fluorescence images of NRK-52E cells. Viable cells were pale green and marked with white arrow, red arrow represents early apoptotic cells (chromatin condensation) were stained bright-green. Late apoptotic cells stained yellow-orange marked by yellow arrow. Cells showing necrosis stained orange-red and marked with blue arrow. Cells treated with serum free media considered as untreated control. Cystone (50 μg/mL) was taken as positive control. Multiple fields were assessed under florescence microscope (Nikon eclipse, Ti) and images were captured at 20X magnification, scale bar 100 μm. Untreated control- Fig.a: AO stained, Fig.b: EB stained and Fig.c: merged image. Oxalate injury—Fig.d: AO stained, Fig.e: EB stained and Fig.f: merged image. AE1 (50 μg/mL) treatment—Fig.g: AO stained, Fig.h: EB stained, Fig.i: merged image. AE2 (50 μg/mL) treatment—Fig.j: AO stained, Fig.k: EB stained, Fig.l: merged image. Cystone (50 μg/mL)—Fig.m: AO stained, Fig.n: EB stained and Fig.o: merged image.