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. 2021 May 3;112(7):2714–2727. doi: 10.1111/cas.14916

FIGURE 3.

FIGURE 3

ZNF213 inhibits Hippo signaling in triple negative breast cancer cells. A, ZNF213 depletion increased the levels of YAP protein in BT549 cells. BT549 cells were transfected with siControl or siZNF213. About 48h later, cells were harvested for western blotting analysis. ZNF213 and YAP protein levels were determined by western blotting.β‐Actin was used as internal reference. B‐C, ZNF213 depletion increased the expression of YAP target gene in BT549 cells. But ZNF213 depletion had no influence on the mRNA expression of YAP. BT549 cells were transfected with siControl or siZNF213. About 48h later, total RNA was extracted for the analysis of gene expression. Each group was repeated three times. ***P < .001 for target gene expression comparison. D, ZNF213 depletion increased the levels of YAP protein in MDA‐MB‐231 cells. MDA‐MB‐231 cells were transfected with siControl or siZNF213. About 48h later, cells were harvested for western blotting analysis. ZNF213 and YAP protein levels were determined by western blotting. β‐Actin was used as internal reference. E‐F, ZNF213 depletion increased the expression of YAP target gene in MDA‐MB‐231 cells. But ZNF213 depletion had no influence on the mRNA expression of YAP. MDA‐MB‐231 cells were transfected with siControl or siZNF213. About 48h later, total RNA was extracted for the analysis of gene expression. Each group was repeated three times. ***P < .001 for target gene expression comparison. G, ZNF213 depletion increased TEAD Luciferase activity in BT549 cells. BT549 cells were transfected with siControl or siZNF213. About 24h later, cells were transfected with the YAP luciferase reporter plasmid and Renilla plasmid. Another 24h later, cells were harvested for the detection of luciferase activity. H, ZNF213 over‐expression decreased the levels of YAP protein in BT549 cells. BT549 cells were transfected with Myc‐ZNF213 or Myc‐tag plasmids. About 48h later, cells were harvested for the analysis of western blotting. Myc‐ZNF213 and YAP protein levels were determined by western blotting.β‐Actin was used as internal reference. I, ZNF213 overexpression decreased the expression of YAP target gene in BT549 cells. BT549 cells were transfected with Myc‐ZNF213 or Myc‐tag plasmids. About 48h later, total RNA was extracted for the analysis of gene expression. Each group was repeated three times. ***P < .001 for target gene expression comparison. J, ZNF213 decreased TEAD Luciferase activity in BT549 cells. BT549 cells were transfected with GFP‐ZNF213 or GFP plasmids. About 24h later, cells were transfected with the YAP luciferase reporter plasmid and Renilla plasmid. Another 24h later, cells were harvested for the detection of luciferase activity