Figure 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.