Fig. 4

Intracellular LOXL1 inhibits the transcriptional activity of YAP. a LOXL1 and YAP constructs alone or combined, as indicated, were transfected into HEK293T cells together with a 3× GTIIC luciferase reporter. The results shown were normalized for transfection efficiency. The cell lysates were immunoblotted with the indicated antibodies (lower panel). Data are shown as the mean ± SD of triplicate independent sets of experiments; statistical significance was assessed by unpaired t-test. *P < 0.05, **P < 0.01. b LOXL1 and YAP alone or combined as indicated were transfected into HEK293T cells for 24 h. Total RNA was prepared, and qRT-PCR was conducted to measure the induction of CDC20, CDX2A, CTGF, and CYR61. Data are shown as the mean ± SD of triplicate independent sets of experiments; statistical significance was assessed by unpaired t-test. ns; non-significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. c LOXL1 or its truncations were transfected into HEK293T cells for 24 h. Flag immunoprecipitates prepared from DMEM and whole cell lysates were analysed by western blotting with a Flag antibody. d In HEK293T cells, LOXL1, LOXL1 △SP, LATS2 and YAP alone or combined as indicated were tested for YAP binding promoter 3× GTIIC luciferase activity induction. The cellular extracts were immunoblotted with the indicated antibodies (lower panel). Data are shown as the mean ± SD of triplicate independent sets of experiments; statistical significance was assessed by unpaired t-test. **P < 0.01, ***P < 0.001. e In HEK293T cells, LOXL1, LOXL1 mutant, LATS2 and YAP alone or combined as indicated were tested for YAP binding promoter 3× GTIIC luciferase activity induction. The cell lysates were immunoblotted with the indicated antibodies (lower panel). The data are shown as the mean ± SD of triplicate independent sets of experiments; ***P < 0.001