Cancer cell adaptation to hypoxia involves a HIF‐GPRC5A‐YAP axis

A

Hypoxia induced increased total YAP expression in a panel of colorectal cancer cell lines. Actin confirmed equal loading; blots are representative of at least two independent experiments. Related to Fig 4.

B–E

qRT–PCR analysis revealed that hypoxia increased the expression of known YAP target genes AREG, BCL2L1 (BCL‐XL), CTGF and CYR61. Representative experiments are shown. Representative examples of n = 3 independent experiments are shown; data are presented as mean ± SD.

F

Increased expression of YAP Ser397 in GPRC5A‐depleted cells was overridden by expression of constitutively active RhoA (G14V) in hypoxia. Expression of dominant negative RhoA (T19N) did not further increase YAP Ser397 phosphorylation in response to GPRC5A depletion.

G

Darker exposure of the GPRC5A blot in Fig 4I, confirming expression of 30–40 kDa and 80 kDa species. Note that YAP siRNA partially diminishes GPRC5A in line with the existence of positive HIF‐GPRC5A‐YAP feedback loop. Asterisk (*) indicates non‐specific band.

Figure EV4. GPRC5A promotes hypoxic cell survival via YAP .