Immunohistochemical staining was performed using a commercial kit, according to the manufacturer’s instructions (ZSGB-BIO, Beijing, China). Primary antibodies used in this study were anti-Apaf-1 (1:200, BA2373), anti-Bax (1:300, BA0315), anti-Bcl-2 (1:200, BA0412) and anti-active caspase-3 (1:300, BA3968), anti-active caspase-9 (1:300, BA0690), all obtained from Boster, Co., Ltd., Beijing, China.
Apaf-1, Bax, Bcl-2, and active caspase-3, active caspase-9 positive signals were observed as brown or yellow granular masses in cells, and their intensities were measured using Motic Med 6.0 CMIAS Image Analysis System (Motic China Group, Co., Ltd., China). A total of 15 fields per gerbil (three fields per section, five sections per gerbil, 400× magnification) were randomly selected and analyzed. The positive staining intensity was calculated as the ratio of the stained area to the total field assessed (Du et al., 2015 (link)).
Apaf-1, Bax, Bcl-2, and active caspase-3, active caspase-9 positive signals were observed as brown or yellow granular masses in cells, and their intensities were measured using Motic Med 6.0 CMIAS Image Analysis System (Motic China Group, Co., Ltd., China). A total of 15 fields per gerbil (three fields per section, five sections per gerbil, 400× magnification) were randomly selected and analyzed. The positive staining intensity was calculated as the ratio of the stained area to the total field assessed (Du et al., 2015 (link)).
Free full text: Click here
