Atorvastatin calcium, Mevalonic acid, GGPP, FPP, Propidium iodide (PI) and 2′,7′-dichlorofluorescein diacetate (DCFH-DA) were purchased from Sigma-Aldrich (St. Louis, MO, USA). 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) was from Amresco (Solon, OH, USA). 5,5′,6,6′-tetrachloro-1, 1′,3,3′-tetraethylbenzimidazole-carbocyanide iodine (JC-1) and cell mitochondria isolation kit were obtained from Beyotime Biotechnology (Shanghai, China). Annexin V-FITC Apoptosis Detection Kit, as well as antibodies against p-pRb (S780) (1:1,000), p27 (1:1,000) and cyclinD1 (1:1,000) were from BD Biosciences Pharmingen (San Jose, CA, USA). Antibodies against pRb (1:1,000), cyclinB1 (1:2,000), cdc2 (1:1,000), caspase-3 (1:1,000), caspase-9 (1:1,000), poly (ADP-ribose) polymerase (PARP) (1:1,000), Cytochrome c (1:1,000), YAP (1:1,000), p-YAP (Ser127) (1:1,000), Rho A (1:1,000), β-actin (1:1,000), anti-mouse (1:2,000), anti-rabbit HRP-conjugated and anti-rabbit Alexa Fluor 488-conjugated secondary antibodies (1:2,000) were from Cell Signaling Technology (Danvers, MA, USA). Antibodies against Bcl-2 (1:1,000), Bax (1:1,000) and Lamin B (1:500) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA).
P yap ser127
Manufactured by Cell Signaling Technology
Sourced in United States
P-YAP(Ser127) is a laboratory reagent used in cell and molecular biology research. It is an antibody that specifically recognizes the phosphorylated form of the Yes-associated protein (YAP) at serine 127 residue. This phosphorylation event is a key regulatory mechanism for the activity and subcellular localization of the YAP transcriptional co-activator.
Automatically generated - may contain errors
Lab products found in correlation
P akt, by Cell Signaling Technology (4 mentions)
Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (4 mentions)
β actin, by Cell Signaling Technology (3 mentions)
Mmp 9, by Cell Signaling Technology (3 mentions)
β actin, by Santa Cruz Biotechnology (3 mentions)
Plats1 thr 1079, by Cell Signaling Technology (2 mentions)
Gapdh, by Cell Signaling Technology (2 mentions)
Protein a g magnetic beads, by Selleck Chemicals (2 mentions)
P erk1 2, by Cell Signaling Technology (2 mentions)
Sds lysis buffer, by Beyotime (2 mentions)
Lats1, by Cell Signaling Technology (2 mentions)
P erk, by Cell Signaling Technology (2 mentions)
P yap ser397, by Cell Signaling Technology (2 mentions)
Yap taz, by Cell Signaling Technology (2 mentions)
Mmp 2, by Cell Signaling Technology (2 mentions)
Histone h3, by Cell Signaling Technology (2 mentions)
Pvdf membrane, by Merck Group (2 mentions)
Gapdh, by Merck Group (2 mentions)
Cytochrome c, by Cell Signaling Technology (1 mentions)
Atorvastatin calcium, by Merck Group (1 mentions)
18 protocols using p yap ser127
1
Atorvastatin Inhibits Proliferation and Induces Apoptosis in Cancer Cells
2
Protein Damage Response Markers Assay
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Anti-γ-H2AX (Ser139), anti-phospho-ATM (Ser 1981), anti-phospho-DNA-PK(Ser2056), anti-phospho-BRCA1 (Ser 1524), anti-Cyr61, anti-survivin,anti-YAP, p-YAP (Ser127) and anti-CyclinB1 were purchased from Cell Signaling Technology (Danvers, MA, USA) or Abcam (Cambridge, MA, USA). Anti-γ-H2AX (Ser139), conjugated with fluorescein isothiocyanate (FITC) was purchased from BD Company (Franklin Lakes, New Jersey, USA). The anti-β-actin antibody, 7-Hydroxystaurosporine (UCN-01) and colcemid were purchased from Sigma-Aldrich (Saint Louis, Missouri, USA), and peroxidase-conjugated goat anti-mouse and goat anti-rabbit secondary antibodies were purchased from ZSGQ-BIOCompany (Beijing, China). 7-Hydroxystaurosporine (UCN-01) and colcemid were purchased from Sigma-Aldrich (Saint Louis, Missouri, USA).
3
Western Blot Analysis of Apoptosis Regulators
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Western blot was performed according to a standard method, as described previously (38 (link)). Antibodies against BAX (#5023), BAD (#9239), BCL2L1 (#2764), BCL2 (#2872), p-MST1 (Thr183)/2(Thr180) (#3681), MST1 (#14946), p-LATS1(Thr1079) (#8654), LATS1 (#9153), p-YAP(Ser127) (#13008), YAP (#14074), NF2 (#6995), and RASSF1 (#86026) were purchased from Cell Signaling Technology, and TAZ from Abcam (ab224239), RASSF5 from Sigma (N5912). The membranes were stripped and reprobed with an anti–α-tubulin antibody (Cell Signaling Technology) as the loading control.
4
Antibody-based Immunoblot Analysis
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Antibodies used for immunoblots were purchased from the indicated companies: p‐YAP (Ser127) (NO.13008), YAP (NO.14074), Lats1 (NO.3477), pLats1 (Ser909) (NO.9157), and pLats1 (Thr1079) (NO.8654) were from (Cell Signalling Technology). SREBP‐1 (Cat#557036), SREBP‐2 (Cat#557037) were from BD Biosciences.
5
Immunoprecipitation Assay for NF2 and YAP Signaling
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For immunoprecipitation assay, artery homogenates or VSMC extracts were prepared in lysis buffer (50 mmol/L Tris·HCl (pH 7.5), 150 mmol/L NaCl, 0.5% IGEPAL CA-630, 0.5% deoxycholic acid, 0.1% SDS, 1 mmol/L EDTA, 1 mmol/L NaF, 0.1 mmol/L Na3VO4, 50 μmol/L phenylmethylsulfonyl fluoride, 5 μg/mL leupeptin, 5 μg/mL aprotinin). Samples were then incubated with primary antibody at 4°C overnight, and immunocomplexes were precipitated after 1 hour of incubation with sepharose A/G beads (Santa Cruz, Delaware, CA). Western blot analysis was performed as described previously [46 (link)]. Extraction of artery and VSMC were lysed with RIPA buffer (Beyotime Institute of Biotechnology, Shanghai, China). Antibodies against phospho (p)-NF2Ser518, NF2, p-YAPSer127, YAP, TEAD1, GAPDH, H3, β-actin were purchased from Cell Signaling Technology (Danvers, MA). The luminescent signal was determined by exposure to x-ray film and quantitative analysis was completed with Image J software (Bethesda, MA).
6
Protein Extraction and Western Blot Analysis
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The cell protein was extracted in SDS lysis buffer (Beyotime, Shanghai, China). The protein samples were electrophoresed on 4–20% polyacrylamide gels (Genshare Biological, China) and transferred to polyvinylidene fluoride (PVDF) membranes (Merck Millipore, USA). The antibodies for PKM2, p-AKT (Ser473), AKT, p-ERK1/2 (Thr202/Tyr204), ERK1/2, Galectin-9, E-cadherin, MMP2, MMP9, p-IKKα/β (Ser176/180), IKKα/β, p-IκBα (Ser32), IκBα, p-NF-κB p65 (Ser536), NF-κB p65, p-AMPK (Thr172), AMPK, p-LATS1 (Thr1079), LATS1, and p-YAP(Ser127) were purchased from Cell Signaling Technology (Danvers, MA, USA). β-actin, α-tubulin and GAPDH antibodies (Proteintech, Rocky Hill, NJ, USA) were used as internal controls. For immunoprecipitation analysis, Cells were lysed with RIPA lysis (Beyotime, Shanghai, China), incubated with specific antibodies and normal IgG and followed by incubation with Protein A/G Magnetic Beads (Bimake, Houston, TX, USA). Immunoreactive bands were visualized with ECL Ultra (New Cell and Molecular Biotech, Suzhou, China).
7
Immunoblotting and Microscopy Protocols
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The following antibodies were purchased from Cell Signaling and used at the indicated dilution for Western blot analysis: Merlin (NF2) (1:1000; 6995), phospho-Merlin Ser518 (1:1000; 9163), YAP (1:1000; 14074), pYAP-Ser127 (1:1000; 4911), LATS1 (1:1000; 3477), phospho-LATS HM (1:1000; 8654), phospho-MK2 Thr334 (1:1000; 3007), phospho-p38 Thr180/Tyr182 (1:1000; 4511), and HA HRP-conjugated (1:10,000; 2999). GAPDH (1:2000) and YAP/TAZ (1:500) were obtained from Santa Cruz Biotechnology (sc-25778 and sc-101199, respectively). Vinculin (1:5000) and Flag HRP conjugated (1:10,000) were from Sigma (V9131 and A8592, respectively. N-Cadherin (1:1000) was from BD Biosciences (610920).
The following antibodies were used for immunofluorescent microscopy experiments at the indicated dilutions: GFP (1:200) was from Abcam (ab6673), PI(4,5)P2 (1:200) was from Echelon Biosciences (Z-G045), Flag (1:500) and HA (1:500) were from Cell Signaling (14793 and 3724, respectively), and YAP (1:200) was from Santa Cruz Biotechnology (sc-101199).
Secondary antibodies Alexa fluor 488 and 555 and phalloidin were from Invitrogen and used in 1:1000 dilution.
Chemicals sorbitol was from Fisher Scientific (S459-500), latrunculin B (Lat B) was from Abcam (ab144291), 2-deoxy-D-glucose (2-DG) was from Sigma (D8375), and p38 inhibitor SB203580 (1202) and JNK inhibitor SP600125 (1496) were from Tocris.
The following antibodies were used for immunofluorescent microscopy experiments at the indicated dilutions: GFP (1:200) was from Abcam (ab6673), PI(4,5)P2 (1:200) was from Echelon Biosciences (Z-G045), Flag (1:500) and HA (1:500) were from Cell Signaling (14793 and 3724, respectively), and YAP (1:200) was from Santa Cruz Biotechnology (sc-101199).
Secondary antibodies Alexa fluor 488 and 555 and phalloidin were from Invitrogen and used in 1:1000 dilution.
Chemicals sorbitol was from Fisher Scientific (S459-500), latrunculin B (Lat B) was from Abcam (ab144291), 2-deoxy-D-glucose (2-DG) was from Sigma (D8375), and p38 inhibitor SB203580 (1202) and JNK inhibitor SP600125 (1496) were from Tocris.
8
Immunostaining Analysis of Neuronal Markers
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After 0.9 and 4% PFA perfusion, fixation, and post-fixation, tissues underwent immunostaining after cryosection (10 μm, embedded in OCT Tissue Tek Medium, Sakura Finetek). After blocking with the appropriate sera, sections were incubated overnight at 4°C with primary antibodies. The next day, sections were washed three times, incubated with appropriate fluorescent secondary antibodies (1:400, molecular probes, Thermo Fisher Scientific Inc., Rockford, IL, United States) 1 h at RT. After DAPI staining, the slides were immediately mounted. Images were taken by SP-8 confocal microscope (Leica, Germany). The primary antibodies used in this work are Tuj1 (1:500, mouse monoclonal; Covance, Cat. MMS435P), SMI-32 (1:400, Abcam, Cat. Ab8135), YAP, and Phosphorylated YAPSer127 (p-YAPSer127) (1:100, Cell Signaling Technology, United States, Cat. 13008).
9
Protein Expression Analysis by Western Blot
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Samples from cultured cells were homogenized in lysis buffer with protease inhibitors, and total protein was extracted. Proteins were mixed with SDS sample buffer and loaded onto 4-12% gradient SDS-PAGE gels. The separated proteins were transferred onto nitrocellulose membranes by the Dry Blotting System (Invitrogen). The membranes were first probed with a specific primary antibody including pYAP (Ser127, Cell Signaling), YAP (Cell Signaling), pERK (Cell Signaling), ERK (Cell Signaling), pAKT (Cell Signaling), AKT (Cell Signaling), p-Smad1/5 (Cell Signaling), Smad1 (Cell Signaling), p-Smad2/3 (Cell Signaling) and GAPDH (Sigma) antibodies and then incubated with an appropriate secondary antibody, followed by visualization with ECL reagents (Thermo, USA).
10
Subcellular Fractionation and Western Blotting
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Nuclear and cytoplasmic fractions were isolated using NE-PER ® Nuclear and Cytoplasmic Extraction Reagents (Thermo Scientific, Rockford, IL, USA). Western blotting was carried out by standard procedures as previously described. [13 (link)] The primary antibodies used for the assay were as follows: anti-human SEMA3F (1:1000) (Chemicon, Temecula, CA, USA), anti-human MST2, MST1, MOB1, p-MOB 1, LATS1/2, YAP/TAZ, cleaved caspase-3, BCL2, p-YAP Ser397, p-YAP Ser127, and CCND2 (1:500) (Cell Signaling Technology, Danvers, MA, USA), anti-human β-actin (1:1000) (Cell Signaling Technology), anti-human CD20 (1:1000) (Abcam Cambridge, MA, USA), anti-human TAZ (1:1000) (Abcam), and anti-human Lamin B1 (1:1000) (Novus Biologicals, Littleton, CO, USA).
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