Selective procedure for the instant identification of cellular apoptosis induced by natural products

Figure 1 Fluorescence microscopy observation of LoVo cell apoptosis. Propidium iodide staining of LoVo cells untreated and treated with 140 µg/mL Pinus massoniana bark extract for 24 hours under fluorescence microscope with G.B. filter. Normal cells depicted in the panel show light and homogeneous staining of their nuclei. In contrast, apoptotic cells represented in the panel show thick and irregular staining of their nuclei as a result of chromatin condensation and nuclear fragmentation. Magnification, 400 ×. N: Normal cells; A: Apoptotic cells; C: Control group; T: Treatment group.

Figure 2 DNA gel electrophoresis of LoVo cell apoptosis. DNA ladder of apoptotic LoVo cells treated with 140 µg/mL Pinus massoniana bark extract for 24 hours. Results of gel electrophoresis of relative low-molecular-weight DNA oozed out of apoptotic LoVo cells. M: DNA marker DL2000; C: Control group; T: Treatment group.

Figure 3 Pathological changes of LoVo cells after treatment with 140 µg/mL Pinus massoniana bark extract for 24 hours or not by flow cytometry analysis. A: Control group, a normal cell cycle distribution of an untreated LoVo cell line stained with propidium iodide (PI), there is no obvious apoptotic curve- sub-G1 curve but with a percentage of 10.9% spontaneous apoptosis. Treatment group, altered cell cycle distribution of a treated LoVo cell line stained with PI, there is an obvious apoptotic curve-sub- G1 curve before G1 curve with a gross percentage of 31.4% induced apoptosis; B: Statistical histogram of LoVo cell apoptosis induced by PMBE treatment compared with Control. ^aP < 0.05 vs Control, n = 3, mean ± SD. C: Control group; T: Treatment group.