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Fig. 4 | Cell Communication and Signaling

Fig. 4

From: Yes-associated protein (YAP) induces a secretome phenotype and transcriptionally regulates plasminogen activator Inhibitor-1 (PAI-1) expression in hepatocarcinogenesis

Fig. 4

YAP transcriptionally regulates PAI-1 expression and extracellular PAI-1 levels. a SiRNA-mediated YAP inhibition in murine Hepa1–6 cells using two independent siRNAs (#1, #2). Analysis was performed 48 h after transfection. Western immunoblot and real-time PCR confirmed efficient YAP silencing. Additional real-time PCR analysis was performed for secreted factors identified after initial expression profiling. b Real-time PCR for YAP and PAI-1 after siRNA-dependent YAP inhibition (Sk-Hep1, left panel). Western immunoblot of total PAI-1 amounts and secreted PAI-1 after YAP inhibition (right panel). The scrambled siRNA transfected control was used for normalization and to calculate relative changes. c Western immunoblot of total YAP and PAI-1 levels in total protein extracts derived from murine WT and YAPS127A transgenic livers followed by densitometric quantification (WT: 1 ± 0.26; YAPS127A: 4.41 ± 1.03, left panel). Detection of secreted PAI-1 in blood plasma samples derived from WT and YAPS127A animals followed by signal quantification (WT: 1 ± 0.16; YAPS127A: 2.95 ± 0.19, right panel). Western immunoblot signals for panel b and c were quantified and normalized to the respective house-keeping genes. ‘scr’ controls represent samples that were transfected with the transfection reagent and non-specific siRNA (40 nM for murine Hepa1–6 cells and 20 nM for human Sk-Hep1 cells) with no similarity to cDNA

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