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. 2023 May 12;30(7):1679–1694. doi: 10.1038/s41418-023-01171-8

Fig. 4. CircRILPL1 activates the Hippo-YAP signaling pathway.

Fig. 4

A The effect of circRILPL1 on the expression level of key proteins of the Hippo signaling pathway was examined by western blotting in NPC cells. B The stability of YAP protein was measured in NPC cells after overexpression of circRILPL1. Cells were treated with cycloheximide (CHX, 50 μg/mL) for 0, 4, 8, 12 h and western blotting was used to measure the expression of YAP protein. The experiments were repeated three times. C The effect of circRILPL1 on the ubiquitination level of YAP protein was detected in NPC cells treated with MG132 for 12 h after overexpression of circRILPL1 plasmid for 48 h. Cell lysates were subjected to immunoprecipitation using anti-YAP antibody followed by western blotting using anti-ubiquitin antibody. D The interaction between YAP and 14-3-3 proteins in NPC cells after overexpression of circRILPL1 was detected by immunoprecipitation using anti-14-3-3 antibody, followed by western blotting using YAP antibody. GAPDH was used as a negative control. E The abundance of YAP protein in nucleus and cytoplasm was examined in NPC cells after overexpression or knockdown of circRILPL1. PARP was used as a nuclear marker and β-tubulin as a cytoplasmic marker. F The result of CTGF luciferase reporter assay showed that circRILPL1 increased the transcriptional activity of YAP in NPC cells. Data were presented as the means ± SD. *p < 0.05, **p < 0.01, ****p < 0.0001.