Skip to main content
. Author manuscript; available in PMC: 2024 May 16.
Published in final edited form as: F S Sci. 2022 Mar 30;3(2):118–129. doi: 10.1016/j.xfss.2022.03.002

FIGURE 4.

FIGURE 4

Effect of A13 on Hippo signaling cascades in COV434 immortalized human granulosa cells. The x-axis is the cell condition of NT (control) or treated with the AKAP13 inhibitor (A13), and the y-axis is the mRNA relative unit fold change. Cells were treated with A13 at 10 or 20 μM for 24 hours. The mRNA and protein levels of several molecules of Hippo signaling by quantitative real-time polymerase chain reaction and Western blot were analyzed. (A) While the mRNA levels of CTGF YAP-responsive gene were unchanged, (B) CTGF protein levels were greatly reduced by A13 treatment. (C and D) BIRC5 and ANKRD1 are YAP-responsive genes that were also reduced by A13 treatment. The levels of (E) YAP1 (a transcriptional effector of Hippo signaling) mRNA and (F) protein were unchanged after A13 treatment at 10 μM compared with untreated control. The (G) mRNA and (H) protein levels of TEAD1 (a binding partner of YAP) were increased after A13 treatment at 10 μM, but (I) the protein levels of TEAD1 appear to be decreased by A13 at 20 μM. The mRNA and protein samples of CTGF, YAP1, and TEAD1 were analyzed with 2 biologic replicates and 3 technical replicates each; those of BIRC5 and ANKRD1 were analyzed with 1 biologic replicate and 3 technical replicates. Error bars = standard error of the mean. For quantitative real-time polymerase chain reaction, a 1-way quantile ANOVA was used for analyses. *P <.05, **P<.01, and ***P<.001. AKAP13 = A-kinase anchoring protein 13; ANKRD1 = ankyrin repeat domain 1;BIRC= baculoviral inhibitors of apoptosis repeat containing; CTGF= connective tissue growth factor; ECM = extracellular matrix; mRNA = messenger ribonucleic acid; NT= not treated; TEAD = Tea domain family; TEAD-luc = TEAD luciferase; YAP = yes-associated protein.