Fig. 5
From: Regulation of Yki/Yap subcellular localization and Hpo signaling by a nuclear kinase PRP4K
PRP4K regulates growth and invasiveness of breast cancer cells. a–b Kaplan–Meier graphs of progression-free survival analysis showing that high YAP expression correlated with poor prognosis (a) whereas high PRP4K (PRPF4B) expression correlated with good prognosis (b) in triple-negative breast cancer patients. Data were obtained from KMPLOTdatabase (http://kmplot.com/). c, d PRP4K depletion decreased Yap phosphorylation (c) and increased YAP protein level (d) in MDA-MB-231 cells. e, f PRP4K depletion in MDAMB231 cells increased Hpo pathway target gene expression (e), which was reversed by Yap RNAi (f). mRNAs of the indicated Hpo target genes were measured by RT-qPCR. Data are means ± s.d. from three independent experiments. * P < 0.05, ** P < 0.01 (Student’s t-test). g PRP4K depletion increased MDA-MB-231 cell proliferation. Cell numbers were measured by the WST-1 assay. Data are means ± s.d. from three independent experiments. ** P < 0.01 (Student’s t-test). NS, not significant. h Wound healing assay of MDA-MB-231 transfected with the indicated siRNA. i Quantification of wound closure at the indicated time points. Data are means ± s.d. from three independent experiments. ** P < 0.01, *** P < 0.001 (Student’s t-test). j Transwell invasion assay of MDAMB-231 transfected with the indicated siRNA. Data are means ± s.d. from three independent experiments. ** P < 0.01, *** P < 0.001 (Student’s t-test). k Western blot analysis of MDA-MB-231 cells infected with the indicated recombinant lentiviruses. l mRNAs of Hpo target genes from MDA-MB-231 cells infected with the indicated lentiviruses were measured by RT-qPCR. Data are means ± s.d. from three independent experiments. ** P < 0.01 (Student’s t-test). m Relative cell numbers of MDA-MB-231 cells infected with the indicated lentiviruses were measured by the WST-1 assay at different time points after infection. Data are means ± s.d. from three independent experiments. *** P < 0.001 (Student’s t-test). n Wound healing assay of MDA-MB-231 infected with the indicated lentiviruses. o Quantification of wound closure of lentiviral infected MDA-MB-231at the indicated time points. Data are means ± s.d. from three independent experiments. ** P < 0.01, *** P < 0.001 (Student’s t-test). Scale bars, 100 μm (h, n) or 20 μm (j)
