Journal: Journal of experimental & clinical cancer research : CR

Article Title: CircGPRC5A enhances colorectal cancer progress by stabilizing PPP1CA and inducing YAP dephosphorylation.

doi: 10.1186/s13046-023-02915-7

Figure Lengend Snippet: Fig. 8 CircGPRC5A can cause dephosphorylation of YAP-ser127 and YAP-ser109 by stabilizing the PPP1CA protein. A Western blotting was used to examine PPP1CA protein after silencing circGPRC5A. B Phosphoproteomics revealed 4,170 phosphopeptides, 4,793 phosphorylation sites, and 2,390 phosphoproteins. C The pie chart illustrates the types of phosphorylated amino acids from quantitative phosphoproteomics analysis. D Upregulated and downregulated protein phosphorylation levels from quantitative phosphoproteomics analysis. E Bubble chart depicting the deltaP_value and phosphorylation site number. F, G Western blotting was used to examine YAP phosphorylation after silencing and overexpressing PPP1CA or circGPRC5A. H–I The effects of YAP dephosphorylation via PPP1CA on circGPRC5A-induced changes were detected by western blotting

Article Snippet: The primary antibodies used were PPP1CA (sc-7482, Santa Cruz Biotechnology), Ub (#3936S CST), UBA1 (ab180125, Abcam), FBXO2 (ab133717, Abcam), YAPSer127 (AP0489, ABclonal), YAP-Ser109 (YP1553, Immunomy), YAP-Ser61(#75784S, CST), YAP (A21216, ABclonal), Pan Phospho-Serine/Threonine (T91067S, Abmart), and GAPDH (10,494–1-AP, Proteintech).

Techniques: De-Phosphorylation Assay, Western Blot, Phospho-proteomics

Journal: Oncogene

Article Title: Expression of the chrXq27.3 miRNA cluster in recurrent ovarian clear cell carcinoma and its impact on cisplatin resistance

doi: 10.1038/s41388-020-01595-3

Figure Lengend Snippet: A Transfection efficacy of two siRNAs for YAP1 (siYAP1 No.1 and No.2). B Cisplatin sensitivity of siYAP1-transfected ES-2 cells measured using MTS assay. The viability of treated cells was compared with that of cells transfected with negative control siRNA (siCtrl) using Student’s t test. C Apoptosis assay of transfected cells treated with 7.5 µM cisplatin. The percentage of apoptotic cells was compared using Student’s t test. D Cisplatin sensitivity of YAP1 inhibitor-treated ES-2 cells measured using MTS assay. The viability of treated cells was compared with that of cells treated with DMSO using Student’s t test. E Expression of miRNAs and YAP1 in YAP1 overexpressed ES-2 cells (FLAG-YAP1) and mock transfected cells (FLAG). The cells were further transfected with combination of miR-509-3p and miR-509-3-5p. F Cisplatin sensitivity of FLAG-YAP1 cells measured using MTS assay. The viability of the two miRNAs transfected cells was compared with that of NC transfected cells using Student’s t test. G Representative images and bar graph of clonogenic assay. The transfected cells were treated with 1 µM cisplatin for 8 days, and the number of colonies was compare using Student’s t test. H The expression of phospho-YAP (Ser127) in the cytoplasm and nucleus of FLAG-YAP1 cells. Error bars represent standard errors of the mean. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: The membranes were blocked in 3% BSA/TBS-T, and the following antibodies were used: PhosphoPlus YAP (Ser127) Antibody Duet #46538 and Anti-rabbit IgG, HRP-linked Antibody #7074 (Cell Signaling Technology, Danvers, MA).

Techniques: Transfection, MTS Assay, Negative Control, Apoptosis Assay, Expressing, Clonogenic Assay