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Fig. 6 | Journal of Experimental & Clinical Cancer Research

Fig. 6

From: Dropwort-induced metabolic reprogramming restrains YAP/TAZ/TEAD oncogenic axis in mesothelioma

Fig. 6

Dropwort extract modulates YAP/TAZ activities. a Histograms showing the 2^Zratio phosphoprotein levels of 12 selected protein known to modulate YAP and TAZ activities. This protein expression was found modulated in MSTO-211H cells treated for 24 h with Fil.v. extract compared to untreated ones (dashed line) (b) Representative protein gel blot of whole cell lysates obtained from MSTO-211H (left side) and MPP-89 (right side) cells treated for 72 h with 50 μg/ml of Fil.v. extract and stained with the indicated antibodies. Actin staining was used as loading control. c Quantitative-PCR. Histograms show the relative level of the indicated genes in MSTO-211H cells treated or not for 72 h with the Fil.v. extract. Bars indicate the average of three independent experiments. Statistics (t-test): p < 0.05. d Upper part: representative protein gel blot of nucleus/cytosol cell lysates obtained from MSTO-211H cells treated as for panel B and stained with the indicated antibodies. Tubulin and H1 staining was used as loading control for cytosol fraction and nuclei fraction respectively. Lower part: quantitative densitometry of YAP (left side) and TAZ (right side) normalized to the loading control. Bars indicate the average of three independent experiments. Statistics (t-test): p < 0.05. e MSTO211-H cells were treated or not with Fil.v. (50 μg/ml), after 24 h cells were transfected with HA-tagged Ubiquitin for further 24 h in combination with MG-132 (25 μM) for 6 h. Protein lysates were then subjected to immunoprecipitation using anti-IgG, anti-Yap or anti-Taz antibodies and subjected to Western Blotting with the indicated antibodies (right side). Arrows indicate ubiquitinated proteins (Ub-HA). Inputs recovered from anti-IgG IPs were loaded onto gels and subjected to immunoblot together with IP samples (left side). f Quantitative densitometry of the abundance of ubiquitinated proteins is reported in (e)

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