Journal: Cellular and Molecular Life Sciences: CMLS

Article Title: Circular RNA circSTX12 regulates osteo-adipogenic balance and proliferation of BMSCs in senile osteoporosis

doi: 10.1007/s00018-025-05684-y

Figure Lengend Snippet: Hippo/YAP signaling mediates circSTX12-related regulation of BMSCs differentiation. A. Western blot analysis of the expression of proteins involved in adipo-osteogenic pathways in circSTX12 knockdown or knockdown rescue experiment in BMSCs after 7 days’ induction in osteogenic medium. n = 3 biological replicates. B . Immunofluorescence analyses of YAP in BMSCs with circSTX12 knockdown or the control. n = 6 biological replicates. YAP (green) and DAPI (blue). Scale bar = 200 μm. C . ARS and ORO staining and quantification assays results on day 14 in circSTX12 knockdown and the control BMSCs after incubated in adipogenic and osteogenic medium containing 2.5 nM YAP inhibitor or not. n = 6 biological replicates. D . Left panel showed the Western blot results of YAP and p-YAP protein levels in BMSCs transfected with ASO-NC, ASO-circSTX12, OE-MST1, or ASO-circSTX12 + OE-MST1. Right panel showed the histogram result in left panel. n = 3 biological replicates. E . ARS and ORO staining and quantification assays results on day 14 in BMSCs transfected with ASO-NC, ASO-circSTX12, OE-MST1, or ASO-circSTX12 + OE-MST1. n = 6 biological replicates. (F) Left panel showed the Western blot results of YAP and p-YAP protein levels in BMSCs transfected with Vector, OE-circSTX12, SI-MST1, or OE-circSTX12 + SI-MST1. Right panel showed the histogram result in left panel. n = 3 biological replicates. (G) ARS and ORO staining and quantification assays results on day 14 in BMSCs transfected with Vector, OE-circSTX12, SI-MST1, or OE-circSTX12 + SI-MST1. Right panel showed the histogram result in left panel. n = 6 biological replicates. All the data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: BMSCs of passge 4 were seeded in 12-well plates at a density of 0.5–0.8 × 10 5 and treated with different concentrations (1.25, 2.5, and 5nM) of YAP Inhibitor (MCE, Italy) dissolved in the osteogenic or adipogenic medium for 14 days.

Techniques: Western Blot, Expressing, Knockdown, Immunofluorescence, Control, Staining, Incubation, Transfection, Plasmid Preparation

Journal: Bioengineered

Article Title: A high level of the long non-coding RNA MCF2L-AS1 is associated with poor prognosis in breast cancer and MCF2L-AS1 activates YAP transcriptional activity to enhance breast cancer proliferation and metastasis

doi: 10.1080/21655979.2022.2074108

Figure Lengend Snippet: MCF2L-AS1 interacts with YAP and thereby modulates its transcriptional regulatory activity . (a). YAP co-precipitation was achieved using 3`-biotinylated synthetic oligonucleotides, with YAP being detected in the resultant eluates via Western blotting. (b). RIP-qPCR was performed to detect the interacting ability of YAP and MCF2L-AS1 among one another. (c). The expression of MCF2L-AS1 was not correlated with YAP in the tumor tissues of BC patients. (d-e). The mRNA and protein level expression of YAP in BC cells was measured following MCF2L-AS1 knockdown. (f). 8 × GTIIC-luciferase activity was assessed in BC cell lines. YAP OE: YAP overexpression; si MCF2L-AS1: MCF2L-AS1 knockdown. (*p < 0.05, **p < 0.01, ***p < 0.001, **** P < 0.0001). (g). ChIP-qPCR analysis shows the occupancy of ANKRD1, CTGF, and CYR61 promoters YAP. Endogenous YAP was immunoprecipitated from control or MCF2L-AS1 silencing BC cells. n = 3 technical replicates per group.

Article Snippet: After cross-linking, 5 μg of the antibody against YAP (Biorbyt, orb193680) was added to immunoprecipitate endogenous YAP.

Techniques: Activity Assay, Western Blot, Expressing, Knockdown, Luciferase, Over Expression, ChIP-qPCR, Immunoprecipitation, Control

Journal: Translational Oncology

Article Title: Piperlongumine suppresses YAP-ET-1-CXCL2 signaling to modulates aggressiveness of triple-negative breast cancer cells

doi: 10.1016/j.tranon.2026.102700

Figure Lengend Snippet: ET-1 is a downstream of YAP and is suppressed by PL. (A) Real-time PCR analysis and Western blotting analysis of ET-1 and YAP expression levels in YAP-knockdown BT549 and MDA-MB-231 cells. (B) qPCR analysis of ET-1 mRNA levels in MDA-MB-231 and BT549 cells treated with PL (5 μ M) for 24 h. (C) Western blotting analysis of YAP, p-YAP (S61), and ET-1 protein levels in BT549 and MDA-MB-231 cells treated with increasing concentrations of PL for 24 h. (D) Representative immunofluorescence images of ET-1 expression in TNBC cells treated with PL (5 μ M) for 24 h. *** p < 0.001 as determined by unpaired t -tests. Scale bar, 50 μ m.

Article Snippet: Antibodies for YAP (#12395), phosphorylated (p)-YAP (S127) (#13008), p-YAP (S61) (#75784), p-AMPK (AMPK; #130429), AMPK (#103487), p-LATS1 (S909) (#9157), and LATS1 (#3477) were purchased from Cell Signaling Technology.

Techniques: Real-time Polymerase Chain Reaction, Western Blot, Expressing, Knockdown, Immunofluorescence

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