bioimagesUnknownYanmei QiThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201111063JCB

Immunostaining showed that HIF-1β (green) was detected in the nucleus of all cells of 3-day embryoid bodies compared to the basement membrane protein perlecan (perl; red). Nuclei were counterstained with DAPI (blue).

Embryoid bodies were cultured for 4, 5 and 10 days and immunostained for the apical polarity marker MUPP1 (green) and basement membrane perlecan (perl; red). Nuclei were counterstained with DAPI (blue).

Immunostaining showed that HIF-2α (green) was localized to the nucleus of the core cells of 3-day embryoid bodies relative to the basement membrane protein perlecan (perl; red). Nuclei were counterstained with DAPI (blue).

Normal embryoid bodies were cultured for 4 days and immunostained for Bim (green) and mitochondrial complex V (COX V; red). Nuclei were counterstained with DAPI (blue). Bim was mainly expressed in epiblast cells and colocalized with COX V.

E5.0 mouse embryos were immunostained for Bnip3 (green) and the basement membrane proteins perlecan (perl; red) and also labeled for nuclei (DAPI; blue).

Five-day AIF y-/ embryoid bodies were immunostained for the apical marker MUPP1 (green) relative to the basement membrane protein perlecan (red). Nuclei were counterstained with DAPI (blue). Apical polarization of the AIF y/- epiblast was not affected despite delayed lumen clearance.

Four-day AIF y/+ embryoid bodies were incubated with 60 nM dihydroethidium (DHE; red) for 30 minutes to detect ROS production and fixed in 3% paraformaldehyde for 10 minutes. Basement membrane was immunostained for anti-laminin α1 (green), and nuclei were counterstained with DAPI (blue). DHE strongly labeled the centrally located cells in AIF y/+ embryoid bodies.

Four-day AIF y/- embryoid bodies were incubated with 60 nM dihydroethidium (DHE; red) for 30 minutes to detect ROS production and fixed in 3% paraformaldehyde for 10 minutes. Basement membrane was immunostained for anti-laminin α1 (green), and nuclei were counterstained with DAPI (blue). DHE fluorescence was much weaker in AIF y/-<br />embryoid bodies relative to AIF y/+.

Live phase micrographs show the differentiation of endoderm and the columnar epiblast epithelium and the formation of a proamniotic-like cavity in 5-day Bnip3 knockdown embryoid bodies. This image was used to quantify cavitation. Bnip3 knockdown significantly delayed the clearance of centrally located cells.

Embryoid bodies were cultured for 4, 5 and 10 days and immunostained for the apical polarity marker MUPP1 (green) and basement membrane perlecan (perl; red). Nuclei were counterstained with DAPI (blue). Bnip3 KD inhibited apoptosis and delayed lumen clearance but had no effect on epiblast polarity.

Live phase micrographs show the differentiation of endoderm and the columnar epiblast epithelium and the formation of a proamniotic-like cavity in 5-day control embryoid bodies. This image was used to quantify cavitation.

Five-day AIF y/+ embryoid bodies were immunostained for the apical marker MUPP1 (green) relative to the basement membrane protein perlecan (red). Nuclei were counterstained with DAPI (blue).

bioimagesImage 136344 (Figure 3 - E)Figure 3 - ESupplemental Figure 2Figure 7 - AImage 136496 (Figure 1 - E)Image 136497 (Figure 2 - E)Image 136509 (Figure 7 - A)Supplemental Figure 2 - AImage 136510 (Figure 7 - A)Image 136503 (Figure 2 - F)Image 136512 (Supplemental Figure 2 - A)Image 136500 (Figure 2 - E)Image 136502 (Figure 2 - F)Figure 1 - EFigure 5 - DFigure 5Figure 2 - EFigure 7Figure 2 - FImage 136506 (Figure 5 - D)Image 136343 (Figure 3 - E)Image 136507 (Figure 5 - D)Figure 1Figure 3Figure 2Yaling HanYanmei QiPeter CarmelietM. Celeste SimonJie LiuXiaoxiang TianJosef M. PenningerShaohua LiAlan M. GrahamfalseBnip3 and AIF cooperate to induce apoptosis and cavitation during epithelial morphogenesis2012-07-02T11:22:41Z2018-11-29T11:22:41Z2018-11-29T11:22:41ZS-JCBD-20111106310.1083/jcb.201111063