Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2021 Aug 26;17(13):3672–3688. doi: 10.7150/ijbs.60110

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The author(s)

This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/). See http://ivyspring.com/terms for full terms and conditions.

PMC Copyright notice

EndoA2 attenuates OGD-induced NRCM apoptosis. (A) CCK-8 results showed that cell number was time-dependently decreased after OGD stimulation (n=6, *p <0.05 vs. control). (B) CCK-8 results showed that cell number was obviously decreased after OGD stimulation for 12 h. EndoA2 overexpression increased cell number, whereas EndoA2 siRNA knockdown further decreased cell number compared with OGD stimulation (n=6, *p <0.05 vs. control, #p <0.05 vs. OGD). (C) Annexin V-FITC/PI flow cytometry analyses of OGD-induced apoptosis in cells with transfection of Ad-EndoA2 or EndoA2 siRNA. In each plot, viable cells are in the lower left quadrant, early apoptotic cells are in the lower right quadrant, and the upper right represents necrotic or late apoptotic cells. (D) Percentage of apoptotic cells was determined by quantitative analyses. EndoA2 overexpression decreased, whereas EndoA2 siRNA knockdown enhanced the apoptotic rate induced by OGD. Ad-lacZ and negative control (neg) had no significant effect on OGD-induced cell apoptosis (n=6, *p <0.05 vs. control, #p <0.05 vs. OGD+neg or OGD+Ad-lacZ). (E) Representative images of TUNEL and DAPI double-staining in cardiomyocytes with different treatments. The first line images represented the DAPI as nuclei and in blue color; the second line images represented the TUNEL as apoptotic nuclei and in red color; the third line images were the combination of the blue and red images. Scale bars: 100 µm. (F) The number of apoptotic nuclei was counted and densitometric analyses showed that OGD stimulation increased the number of apoptotic nuclei, which was decreased by EndoA2 overexpression, but further increased by EndoA2 siRNA knockdown (n=6, **p <0.01 vs. control, ##p <0.01 vs. OGD+neg or OGD+Ad-lacZ). (G-J) The effects of EndoA2 overexpression or siRNA knockdown on OGD-induced Bax, Bcl-2 and cleaved caspase-3 expression. Densitometric analyses showed that OGD stimulation decreased the expression of Bcl-2 and the ratio of Bcl-2/Bax, whereas increased the expression of Bax and cleaved caspase-3. Overexpression of EndoA2 abrogated the effects of OGD on these proteins, whereas silence of EndoA2 enhanced them (n=6, *p <0.05 vs. control, ##p <0.01 vs. OGD).

EndoA2 attenuates OGD-induced NRCM apoptosis. (A) CCK-8 results showed that cell number was time-dependently decreased after OGD stimulation (n=6, *p <0.05 vs. control). (B) CCK-8 results showed that cell number was obviously decreased after OGD stimulation for 12 h. EndoA2 overexpression increased cell number, whereas EndoA2 siRNA knockdown further decreased cell number compared with OGD stimulation (n=6, *p <0.05 vs. control, #p <0.05 vs. OGD). (C) Annexin V-FITC/PI flow cytometry analyses of OGD-induced apoptosis in cells with transfection of Ad-EndoA2 or EndoA2 siRNA. In each plot, viable cells are in the lower left quadrant, early apoptotic cells are in the lower right quadrant, and the upper right represents necrotic or late apoptotic cells. (D) Percentage of apoptotic cells was determined by quantitative analyses. EndoA2 overexpression decreased, whereas EndoA2 siRNA knockdown enhanced the apoptotic rate induced by OGD. Ad-lacZ and negative control (neg) had no significant effect on OGD-induced cell apoptosis (n=6, *p <0.05 vs. control, #p <0.05 vs. OGD+neg or OGD+Ad-lacZ). (E) Representative images of TUNEL and DAPI double-staining in cardiomyocytes with different treatments. The first line images represented the DAPI as nuclei and in blue color; the second line images represented the TUNEL as apoptotic nuclei and in red color; the third line images were the combination of the blue and red images. Scale bars: 100 µm. (F) The number of apoptotic nuclei was counted and densitometric analyses showed that OGD stimulation increased the number of apoptotic nuclei, which was decreased by EndoA2 overexpression, but further increased by EndoA2 siRNA knockdown (n=6, **p <0.01 vs. control, ##p <0.01 vs. OGD+neg or OGD+Ad-lacZ). (G-J) The effects of EndoA2 overexpression or siRNA knockdown on OGD-induced Bax, Bcl-2 and cleaved caspase-3 expression. Densitometric analyses showed that OGD stimulation decreased the expression of Bcl-2 and the ratio of Bcl-2/Bax, whereas increased the expression of Bax and cleaved caspase-3. Overexpression of EndoA2 abrogated the effects of OGD on these proteins, whereas silence of EndoA2 enhanced them (n=6, *p <0.05 vs. control, ##p <0.01 vs. OGD).