polyclonal antibodies against phospho yap ser127  (Bioss)

Journal: EBioMedicine

Article Title: The cellular prion protein controls the mesenchymal-like molecular subtype and predicts disease outcome in colorectal cancer

doi: 10.1016/j.ebiom.2019.07.036

Figure Lengend Snippet: PrP C controls C4/CMS4 genes via YAP/TAZ. (a) GSEA analysis showing enrichment of the Cordenonsi YAP conserved signature in the genes most correlated with PRNP expression in the Marisa study. (b) Significance expressed as –Log ( p -values) of the enrichment for the regulation of the Cordenonsi YAP conserved signature and the YAP1 up pathway in the genes most correlated to PRNP gene expression in the TCGA colon, Marisa and Medico studies. (c) Analysis of the distribution of the correlation indexes between protein levels and mRNA expression of PRNP expression in the TCGA colon study shows enrichment of YAP and TAZ. Pearson's correlation coefficient R = 0.25 for YAP and TAZ, p < .00001. (d) Analysis of gene expression of a panel of YAP/TAZ targets ( CYR61 , CTGF and AXL ) in the Marisa study demonstrates significant correlations with mRNA expression of PRNP . (e) Correlations indexes (R, Pearson) between mRNA expression of PRNP and YAP1 , WWTR1 , CYR61 , CTGF and AXL in the TCGA colon, Marisa and Medico studies. (f) Western blot analysis of the expression of TAZ, phospho-S127-YAP and total YAP in PRNP -silenced vs. control MDST8 cells. (g) qRT-PCR analysis of the expression of CYR61 , CTGF , AMOTL2 , AXL and GAS6 in PRNP -silenced vs. control MDST8 cells. (h) qRT-PCR analysis of the expression of C4/CMS4 genes in MDST8 cells transfected with siRNA against YAP or TAZ vs. control MDST8 cells. Results are expressed as means of n = 2 independent triplicates of cell preparations ± s.e.m. * p < .05, ** p < .01, *** p < .001 vs. control (Student's t -test).

Article Snippet: Polyclonal rabbit antibodies against phospho-YAP (S127) were from Cell Signalling Technology (Danvers, MA, USA).

Techniques: Expressing, Gene Expression, Western Blot, Control, Quantitative RT-PCR, Transfection

Journal: Scientific reports

Article Title: Plasticity of the inner cell mass in mouse blastocyst is restricted by the activity of FGF/MAPK pathway.

doi: 10.1038/s41598-017-15427-0

Figure Lengend Snippet: Figure 5. Effect of FGF/MAPK inhibition on isolated E3.5 ICMs. (A) Control E3.5 ICM cultured 48 hrs, (B–D) ICMs derived from E3.5 blastocysts and afterwards cultured 48 hr hrs in 2inh: (B) ICM with the outer PE layer, (C) ICM with the outer TE layer, and (D) ICM with outer heterogeneous (PE and TE) layer, (E,F) ICMs derived from E3.5 blastocysts pre-incubated in 2inh. media from the 8-cell stage: (E) ICM with the outer TE layer, (F) Blastocyst with the TE, EPI and PE, (G) The time schedule of inhibitor treatment; red and black arrows indicate the culture periods in the presence or absence of inhibitors, respectively, (H) Composition of epithelial layer in isolated ICMs cultured 48 hrs, (I,J) Polarisation of E3.5 ICMs (isolated from embryos pre-incubated in 2inh. conditions from the 8-cell stage) immediately (I) and 24 hrs after IS (J), (K,L) Localisation of pYAP in E3.5 ICMs (isolated from embryos pre-incubated in 2inh. conditions from the 8-cell stage) immediately (K) and 24 hrs after IS (L), (M,N) Localisation of nuclear YAP in E3.5 ICMs immediately (M) and 24 hrs after IS (N); yellow (*) indicates blastocyst cavity, orange arrows indicate ICM cells devoid of pYAP signal. Scale: 20 μm.

Article Snippet: Primary antibodies used were: rabbit polyclonal antibody against Gata4 (1:100, Santa Cruz, #sc-9053), goat polyclonal antibody against Sox17 (1:100, R&D Systems, #AF1924), mouse monoclonal antibody against Oct4 (1:100, Santa Cruz, #sc-5279), rabbit polyclonal antibody against Nanog (1:100, ReproCELL, #RCAB002P-F), rabbit polyclonal antibody against Sox2 (1:100, Abcam, #ab97959), mouse monoclonal antibody against Cdx2 (1:50, BioGenex, #MU392A-UC), mouse monoclonal antibody against YAP1 (1:100, Abcam, #ab56701), rabbit polyclonal antibody against pYAP (1:100, Cell Signaling Technology, #4911), rabbit monoclonal antibody against pERM (1:400, Cell Signaling Technology, #3141).

Techniques: Inhibition, Isolation, Control, Cell Culture, Derivative Assay, Incubation