rabbit anti phospho yap1  (Proteintech)

Journal: Cellular & Molecular Biology Letters

Article Title: CDH4/UBA1/RBMX axis promotes polycystic ovary syndrome progression through YAP1 activation

doi: 10.1186/s11658-026-00900-4

Figure Lengend Snippet: Mechanism through which CDH4 regulates YAP1. A The mass spectrum of CDH4 specifically binds the first ten molecules of the protein. B Co-IP of CDH4 and RBMX. C Fluorescence co-localization of CDH4 and RBMX with scale bar of 20 μM. D The protein level of RBMX after knocking down CDH4. E RBMX protein expression after knockdown. F Rbmx mRNA expression after knockdown, N = 4. G The mRNA of Yap1 after knocking down RBMX, N = 4. H Yap1 mRNA stability assay. Cells were transfected with siRBMX or siNC, followed by treatment with actinomycin D (5 µg/mL). Yap1 mRNA levels were quantified by qRT-PCR at the indicated times (0, 1, 2, 4, 6, and 8 h) post-treatment. We normalized the mRNA level at the 0-h time point to 1 for each group (siNC and siRBMX), using it as the baseline, N = 3. I YAP1 protein levels after RBMX knockdown. J Fluorescence intensity of YAP1 after knocking down RBMX. Scale, 50 μM N = 3

Article Snippet: The following antibodies were employed: CDH4 (1:500; Santa Cruz, sc-398306), YAP1 (1:500; Santa Cruz, sc-376830), Phospho-YAP1-S127 (1:500; ABclonal, AP0489), RBMX (1:2000; ABclonal, A21963), UBA1 (1:2000; ABclonal, A9254), Ubiquitin (1:2000; Proteintech, 10201-2-AP), Bcl-2 (1:2000; ABclonal, A19693), Caspase-3 (1:2000; ABclonal, A2156), IL-6 (1:1000; Proteintech, 66146-1-Ig), TNF-α (1:1000; Proteintech, 60291-1-Ig), and GAPDH (1:50,000, Proteintech, 60004-1-Ig).

Techniques: Co-Immunoprecipitation Assay, Fluorescence, Expressing, Knockdown, Stability Assay, Transfection, Quantitative RT-PCR

Journal: Cellular & Molecular Biology Letters

Article Title: CDH4/UBA1/RBMX axis promotes polycystic ovary syndrome progression through YAP1 activation

doi: 10.1186/s11658-026-00900-4

Figure Lengend Snippet: RBMX-dependent regulation of YAP1 by CDH4. A Colony formation assays for indicated cells. B CCK-8 assay for indicated cells, N = 4. C Protein expression levels of different molecules under 20 μM DHEA culture for 2 days. D , E Mito-Tracker staining in NC and RBMX-knockdown cells treated with 20 μM DHEA for 2 days. Scale, 100 μM, N = 3. F CCK-8 assay for functional recovery experiments, N = 4. G Protein expression levels of different molecules in functional recovery experiments under 20 μM DHEA culture for 2 days. H , I Representative images and quantification of Mito-Tracker staining for different groups under 20 μM DHEA for 2 days. Scale, 100 μM, N = 3

Article Snippet: The following antibodies were employed: CDH4 (1:500; Santa Cruz, sc-398306), YAP1 (1:500; Santa Cruz, sc-376830), Phospho-YAP1-S127 (1:500; ABclonal, AP0489), RBMX (1:2000; ABclonal, A21963), UBA1 (1:2000; ABclonal, A9254), Ubiquitin (1:2000; Proteintech, 10201-2-AP), Bcl-2 (1:2000; ABclonal, A19693), Caspase-3 (1:2000; ABclonal, A2156), IL-6 (1:1000; Proteintech, 66146-1-Ig), TNF-α (1:1000; Proteintech, 60291-1-Ig), and GAPDH (1:50,000, Proteintech, 60004-1-Ig).

Techniques: CCK-8 Assay, Expressing, Staining, Knockdown, Functional Assay

Journal: Cellular & Molecular Biology Letters

Article Title: CDH4/UBA1/RBMX axis promotes polycystic ovary syndrome progression through YAP1 activation

doi: 10.1186/s11658-026-00900-4

Figure Lengend Snippet: CDH4/UBA1/RBMX complex mediates the ubiquitination and proteasomal degradation of RBMX in KGN cells. A The mRNA level of Rbmx after knocking down CDH4, N = 4. B The protein level of RBMX after CDH4 was knocked down after cycloheximide was added at different times. C RBMX protein levels in different groups with MG132 and chloroquine. D Ubiquitination status of RBMX upon CDH4 knockdown assessed by IP. E Co-IP experiments of UBA1 with RBMX and CDH4, respectively. F IP experiment with RBMX antibody pull-down after knocking down CDH4. G The mRNA expression of Uba1 after overexpressing, N = 4. H The protein expression of UBA1 after overexpressing. I The protein expression levels of UBA1 and RBMX in overexpressed UBA1 and added MG132 groups. J The UBA1 and RBMX protein levels following CHX chase in UBA1-overexpressing cells. K The protein expression level of UBA1 after knocking down CDH4

Article Snippet: The following antibodies were employed: CDH4 (1:500; Santa Cruz, sc-398306), YAP1 (1:500; Santa Cruz, sc-376830), Phospho-YAP1-S127 (1:500; ABclonal, AP0489), RBMX (1:2000; ABclonal, A21963), UBA1 (1:2000; ABclonal, A9254), Ubiquitin (1:2000; Proteintech, 10201-2-AP), Bcl-2 (1:2000; ABclonal, A19693), Caspase-3 (1:2000; ABclonal, A2156), IL-6 (1:1000; Proteintech, 66146-1-Ig), TNF-α (1:1000; Proteintech, 60291-1-Ig), and GAPDH (1:50,000, Proteintech, 60004-1-Ig).

Techniques: Ubiquitin Proteomics, Knockdown, Co-Immunoprecipitation Assay, Expressing

Journal: Journal of Translational Medicine

Article Title: TROP2 confers resistance to oxidative stress-induced cancer cell death through YAP/HMOX1 signaling

doi: 10.1186/s12967-026-07955-z

Figure Lengend Snippet: TROP2 suppression sensitized NSCLC to RSL3-induced ferroptosis in vivo. ( A ) Schematic diagram of the in vivo experimental timeline. Nude mice were subcutaneously injected with control (shCON) or TROP2-knockdown (shTROP2) PC9 cells, followed by intraperitoneal injection of RSL3 (5 mg/kg) or vehicle every other day. ( B ) Representative photographs of dissected subcutaneous tumors from each treatment group at the endpoint of the study. ( C ) Final tumor weights from each group at the endpoint. Data are presented as mean ± SD ( n = 6). ( D ) Tumor growth curves showing tumor volume over time for the four experimental groups. Data are presented as mean ± SEM ( n = 6 mice per group). ( E ) Representative IHC images of tumor sections stained for TROP2, YAP1, HMOX1, ACSL4, and SLC7A11 from the indicated groups. Scale bar, 100 μm. Data are presented as mean ± SD ( n = 6, C) or mean ± SEM ( n = 6 mice per group, D ). Statistical significance for differences among groups was determined by one-way ANOVA followed by Tukey’s post hoc test (C, and for the final time point in D). ∗ P < 0.05, ∗∗ P < 0.01

Article Snippet: After that, the membranes were blocked in 5% fat-free milk for 1 h. The membranes were incubated with the primary antibodies including TROP2 (1:1000, cat #47866, CST), GPX4 (1:1000, cat #AF7020, Beyotime), SLC7A11 (1:1000, cat #AF7992, Beyotime), ACSL4 (1:1000, cat #AG1908, Beyotime), HMOX1 (1:1000, cat #AG2181, Beyotime), YAP1 (1:1000, cat #14074, CST), P-YAP1 (1:1000, cat #13619, CST), LATS1 (1:1000, cat #3477, CST), p-LATS1 (1:1000, cat #9157, CST), Histone3 (1:1000, cat #9715, CST),α-Tubulin (1:1000, cat #2144, CST) at 4 °C overnight and the secondary antibodies at room temperature for 2 h. Protein bands were visualized using the ECL (Bio-Rad, Hercules, CA, USA).

Techniques: In Vivo, Injection, Control, Knockdown, Staining

Journal: Journal of Translational Medicine

Article Title: TROP2 confers resistance to oxidative stress-induced cancer cell death through YAP/HMOX1 signaling

doi: 10.1186/s12967-026-07955-z

Figure Lengend Snippet: TROP2 confers ferroptosis resistance by inactivating YAP to suppress HMOX1 transcription. ( A ) YAP1 mRNA levels in H292 and PC9 cells with TROP2 knockdown, as determined by qRT-PCR. ( B ) YAP1 mRNA levels in PC9 and A549 cells with TROP2 overexpression, as determined by qRT-PCR. ( C ) Representative IF images showing enhanced nuclear localization of YAP (green) in PC9 cells upon TROP2 knockdown. Nuclei were stained with DAPI (blue). Scale bar, 50 μm. ( D ) Representative IF images showing suppressed nuclear localization of YAP (green) in PC9 cells upon TROP2 overexpression. Scale bar, 50 μm. ( E ) Western blot analysis of YAP protein levels in nuclear and cytoplasmic fractions of PC9 and H292 cells with or without TROP2 knockdown. ( F ) Quantitative analysis (bar graph) of the bands was shown adjacent to the representative blots (from E). ( G, H ) Cell viability assessed by CCK-8 assay in PC9 and H292 cells with TROP2 knockdown, with or without concomitant YAP inhibition, following RSL3 treatment. ( I,J ) Measurement of cellular GSH levels in PC9 and H292 cells under the indicated conditions. ( K, L ) Measurement of cellular MDA levels in PC9 and H292 cells under the indicated conditions. Data are presented as mean ± SD ( n = 3). Statistical significance was determined by two-tailed unpaired Student’s t-test ( A , B , F ); two-way ANOVA followed by Sidak’s post hoc test or three-way ANOVA followed by Sidak’s post hoc test ( G - L ). ∗ P < 0.05, ∗∗ P < 0.01

Article Snippet: After that, the membranes were blocked in 5% fat-free milk for 1 h. The membranes were incubated with the primary antibodies including TROP2 (1:1000, cat #47866, CST), GPX4 (1:1000, cat #AF7020, Beyotime), SLC7A11 (1:1000, cat #AF7992, Beyotime), ACSL4 (1:1000, cat #AG1908, Beyotime), HMOX1 (1:1000, cat #AG2181, Beyotime), YAP1 (1:1000, cat #14074, CST), P-YAP1 (1:1000, cat #13619, CST), LATS1 (1:1000, cat #3477, CST), p-LATS1 (1:1000, cat #9157, CST), Histone3 (1:1000, cat #9715, CST),α-Tubulin (1:1000, cat #2144, CST) at 4 °C overnight and the secondary antibodies at room temperature for 2 h. Protein bands were visualized using the ECL (Bio-Rad, Hercules, CA, USA).

Techniques: Knockdown, Quantitative RT-PCR, Over Expression, Staining, Western Blot, CCK-8 Assay, Inhibition, Two Tailed Test

Journal: PLOS One

Article Title: TL1A serves as a positive regulator to promote adipocyte differentiation

doi: 10.1371/journal.pone.0343036

Figure Lengend Snippet: MEFs were treated with standard differentiation medium and TL1A at the specified concentrations (0, 10, 20, 50, 100, 200 ng/mL) for 7 days. (A) Total cellular proteins were extracted, and the expression levels of β-catenin, phosphorylated YAP1 S127 (p-YAP1 S127 ) and total YAP1 proteins were determined by Western blotting. (B) The relative amounts of β-catenin protein expression were calculated according to the grayscale values and were showed in the histogram. (C) The quantitation of ratio (p-YAP1 S127 /total YAP1) to reflect the inactivation of YAP1 signaling pathway. (D-E) At the end of adipogenesis induction, protein expression of phosphorylated β-catenin (p- β-catenin) and YAP1 in the cytoplasm extract from MEFs was determined by Western blotting. (F-G) Nuclear extracts were isolated from the MEFs and analyzed for protein expression by Western blotting. Nuclear proteins were isolated to evaluated the expression of β-catenin, YAP1 and PPARγ proteins through Western blotting, accompanied by a quantitative analysis of band density. GAPDH and Lamin A/C served as the internal controls for cytoplasmic and nuclear extract, respectively. All the histograms represent the relative expression levels of proteins normalized by GAPDH or Lamin A/C. * P <0.05, ** P <0.01, *** P <0.001 vs . the group of MEFs without TL1A treatment.

Article Snippet: Rabbit anti-C/EBPα (Cat# A0904), C/EBPβ (Cat# A0711), β-catenin (Cat# A11932), phospho-YAP1-S127 (Cat# AP0489), YAP1 (Cat# A1002) polyclonal antibodies were purchased from ABclonal Technology (Wuhan, Hubei, China).

Techniques: Expressing, Western Blot, Quantitation Assay, Isolation

Journal: PLOS One

Article Title: TL1A serves as a positive regulator to promote adipocyte differentiation

doi: 10.1371/journal.pone.0343036

Figure Lengend Snippet: Exogenous TL1A triggers the phosphorylation of YAP1 at Ser127, resulting in its cytoplasmic retention and functional inactivation. This event subsequently reduces β-catenin stability and prevents its nuclear translocation, thereby triggering the expression of essential adipogenic regulators and lipid metabolism-related proteins. Additionally, TL1A-mediated regulation of ABCA1 and ABCG1 expression facilitates cholesterol homeostasis, providing essential substrates for lipid droplet biogenesis. Collectively, these molecular mechanisms orchestrate the differentiation of MEFs and 3T3-L1 preadipocytes into terminally differentiated adipocytes.

Article Snippet: Rabbit anti-C/EBPα (Cat# A0904), C/EBPβ (Cat# A0711), β-catenin (Cat# A11932), phospho-YAP1-S127 (Cat# AP0489), YAP1 (Cat# A1002) polyclonal antibodies were purchased from ABclonal Technology (Wuhan, Hubei, China).

Techniques: Phospho-proteomics, Functional Assay, Translocation Assay, Expressing