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. Author manuscript; available in PMC: 2015 Apr 20.
Published in final edited form as: Oncogene. 2012 Apr 23;32(10):1266–1273. doi: 10.1038/onc.2012.147

Figure 5.

Figure 5

Knockdown of PTPN14 induces YAP nuclear retention and increases cell migration in a YAP-dependent manner. (a) Knockdown of PTPN14 decreases the level of phospho-Ser127 YAP. Immunoblot demonstrates efficient knockdown of PTPN14 with the two independent lentiviral shPTPN14 constructs in MCF10A cells. Reduced phospho-Ser127 YAP level was detected in PTPN 14-knockdown cells compared with the control cells, with no change observed at the total YAP protein level. β-actin used as loading control. (b) Knockdown of PTPN14 results in YAP nuclear retention at high cell density in MCF10A cells. At high cell density, immunofluorescence microscopy shows that YAP (magnification, ×63) was localized in the cytoplasm of vector control cells, whereas YAP nuclear retention remained in the PTPN14-knockdown cells. Quantification of YAP localization in either nuclear or cytoplasm were indicated. (c) qRT–PCR analysis of the YAP target genes in MCF10A cells. GAPDH was used as an internal control. Error bars equal ± s.d. (d) Knockdown of PTPN14 increases YAP-dependent cell migration. Control, two independent PTPN14-knockdown or YAP and PTPN14 concomitant knockdown MCF10A cells were plated onto 8-µm transwell filters and allowed to migrate for 24 h. Data are the mean number of migrated cells per field of four fields from each of the triplicate wells. Error bars equal ± s.d. of three independent experiments.