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A
Overexpression of YAP induced the transcription of BCAR4. The transcription of BCAR4 and YAP downstream genes was examined in empty vector (EV) and YAP‐overexpressing MCF10A cells (mean ± s.d., n = 3 biological replicates, Student's t‐test).
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B
The transcriptional activity of YAP was required for the upregulation of BCAR4. The transcription of BCAR4 was examined by real‐time PCR in HEK293A cells stably expressing indicated YAP mutants (mean ± s.d., n = 3 biological replicates, Student's t‐test).
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C
Glucose induced the expression of BCAR4. HEK293A cells were glucose‐starved for 24 h and then stimulated with glucose (25 mM) for the indicated intervals (mean ± s.d., n = 3 biological replicates, Student's t‐test).
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D
Two YAP/TEAD‐binding elements were identified in the BCAR4 promoter region.
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E, F
YAP/TEAD directly regulates the transcription of BCAR4. Chromatin immunoprecipitation assay was performed by using YAP antibody in the glucose‐starved or stimulated MDA‐MB‐231 (E). BCAR4 promoter luciferase reporter assay was performed by overexpressing YAP‐5SA and TEAD4 in HEK293T (F) (mean ± s.d., n = 3 biological replicates, Student's t‐test).
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G
Distribution of alteration frequency of
BCAR4 in multiple cancer types. Colors indicate different cancer types. Details in parentheses indicate the source of the corresponding tumor dataset.
BCAR4 was analyzed in multiple cancers by using public database cBioportal (
http://www.cbioportal.org).
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H
The expressions of BCAR4 and YAP are positively correlated in breast cancer. RNAScope® detection of BCAR4 and immunohistochemical staining of YAP were performed by using breast cancer tissue arrays. Brown staining indicates positive immune reactivity. The region in each box is enlarged below. Scale bar, 200 μm. Correlations between YAP and BCAR4 levels in human breast tumors were analyzed as a table. Statistical significance was determined by the chi‐square test; R, correlation coefficient.
