ZFIN Image: Greenhough et al., 2018, 3

Image

Figure Caption

3 GPRC5A promotes hypoxic cancer cell survival

A

GPRC5A depletion markedly increases caspase‐3 activation/PARP cleavage during hypoxia.

B

Three independent siRNA sequences targeting GPRC5A induce caspase‐3 activation/PARP cleavage during hypoxia.

C

GPRC5A depletion reduces hypoxic cell growth/survival. Crystal violet cell assays show reduced cell growth/survival in GPRC5A‐depleted cells during hypoxia (n = 3).

D

Expression of an siRNA‐resistant GPRC5A cDNA rescues hypoxic GPRC5A‐depleted cells from apoptosis. Upper: doxycycline‐induced expression of GPRC5A^si1R rescues increased caspase‐3/PARP cleavage induced by GPRC5A depletion in hypoxia. Lower: generation of an siRNA1‐resistant GPRC5A cDNA by synonymous mutations.

E

GPRC5A depletion in hypoxia induces apoptosis as determined by the violet ratiometric membrane asymmetry probe/dead cell apoptosis assay and flow cytometry (n = 3 independent experiments).

F

Caspase inhibitor QVD prevented caspase‐3 activation/PARP cleavage by GPRC5A depletion in hypoxia.

Data information: Asterisks (*) indicate non‐specific band. Level adjustments were made to images in Adobe Photoshop post‐acquisition for clarity (equal changes applied to the entire image). Representative examples of n = 3 independent experiments are shown; data are presented as mean ± SEM. One‐way ANOVA with Tukey's multiple comparisons test was carried out in (C and F).Source data are available online for this figure.

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ EMBO Mol. Med.