bs-3504R Search Results


phospho mst1  (Bioss)
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GPR137 suppresses Hippo signaling by MST. A Protein levels of <t>pMST1,</t> <t>MST1,</t> pLATS1, LATS1, pLATS2 and LATS2 in AGS cells transfected with GPR137 (GPR137, +) or an empty vector (GPR137, -) for 24 h. One representative of 3 independent blots is shown. B Protein levels of pTAZ, TAZ, pYAP, YAP, CTGF and CYR61 in AGS cells transfected with GPR137 (GPR137, +) or an empty vector (GPR137, -) for 24 h. One representative of 3 independent blots is shown. C Co-immunoprecipitation of HA-tagged GPR137 and Flag-tagged MST1 in HEK293T cells. IP: Flag; WB: HA. N = 3. D Co-immunoprecipitation of endogenous GPR137 and MST1 in AGS cells. IP: MST1; WB: GPR137. IgG was used as a negative control. One representative of 3 independent blots is shown. E Co-immunoprecipitation of endogenous MST1 and LATS1 in the presence of GPR137 in AGS cells. IP: MST1; WB: LATS1. IgG was used as a negative control. One representative of 3 independent blots is shown. F Co-immunoprecipitation of endogenous MST1 and LATS1 in the presence of XMU-MP-1 in AGS cells. IP: MST1; WB: LATS1. IgG was used as a negative control. One representative of 3 independent blots is shown. G Protein expression examination of MST1 and MST2 in MST1/2 -knockout ( MST1/2 -KO) AGS cells or control AGS cells. One representative of 3 independent blots is shown. H Nucleo-cytoplasmic separation assays in MST1/2 -knockout AGS ( MST1/2 -KO) cells or control cells, and protein levels of YAP and TAZ were detected. Proteins in nucleus (left) and in cytoplasm (right). One representative of 3 independent blots is shown. I Nucleo-cytoplasmic separation assays in GPR137-transfected AGS (GPR137, +) cells or control cells (GPR137, -), and protein levels of YAP and TAZ were detected. Proteins in nucleus (upper) and in cytoplasm (lower). One representative of 3 independent blots is shown. J TEAD-luciferase assays in MST1/2 -knockout ( MST1/2 -KO) AGS cells or control AGS cells transfected with GPR137 or an empty vector (control). ** p < 0.01; error bar, SD. N = 3. K TEAD-luciferase assays in MST1/2 -knockout ( MST1/2 -KO) AGS cells or control AGS cells transfected with GPR137 shRNA or control scrambled shRNA. ** p < 0.01; error bar, SD. N = 3
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GPR137 suppresses Hippo signaling by MST. A Protein levels of pMST1, MST1, pLATS1, LATS1, pLATS2 and LATS2 in AGS cells transfected with GPR137 (GPR137, +) or an empty vector (GPR137, -) for 24 h. One representative of 3 independent blots is shown. B Protein levels of pTAZ, TAZ, pYAP, YAP, CTGF and CYR61 in AGS cells transfected with GPR137 (GPR137, +) or an empty vector (GPR137, -) for 24 h. One representative of 3 independent blots is shown. C Co-immunoprecipitation of HA-tagged GPR137 and Flag-tagged MST1 in HEK293T cells. IP: Flag; WB: HA. N = 3. D Co-immunoprecipitation of endogenous GPR137 and MST1 in AGS cells. IP: MST1; WB: GPR137. IgG was used as a negative control. One representative of 3 independent blots is shown. E Co-immunoprecipitation of endogenous MST1 and LATS1 in the presence of GPR137 in AGS cells. IP: MST1; WB: LATS1. IgG was used as a negative control. One representative of 3 independent blots is shown. F Co-immunoprecipitation of endogenous MST1 and LATS1 in the presence of XMU-MP-1 in AGS cells. IP: MST1; WB: LATS1. IgG was used as a negative control. One representative of 3 independent blots is shown. G Protein expression examination of MST1 and MST2 in MST1/2 -knockout ( MST1/2 -KO) AGS cells or control AGS cells. One representative of 3 independent blots is shown. H Nucleo-cytoplasmic separation assays in MST1/2 -knockout AGS ( MST1/2 -KO) cells or control cells, and protein levels of YAP and TAZ were detected. Proteins in nucleus (left) and in cytoplasm (right). One representative of 3 independent blots is shown. I Nucleo-cytoplasmic separation assays in GPR137-transfected AGS (GPR137, +) cells or control cells (GPR137, -), and protein levels of YAP and TAZ were detected. Proteins in nucleus (upper) and in cytoplasm (lower). One representative of 3 independent blots is shown. J TEAD-luciferase assays in MST1/2 -knockout ( MST1/2 -KO) AGS cells or control AGS cells transfected with GPR137 or an empty vector (control). ** p < 0.01; error bar, SD. N = 3. K TEAD-luciferase assays in MST1/2 -knockout ( MST1/2 -KO) AGS cells or control AGS cells transfected with GPR137 shRNA or control scrambled shRNA. ** p < 0.01; error bar, SD. N = 3

Journal: Biology Direct

Article Title: GPR137 inactivates Hippo signaling to promote gastric cancer cell malignancy

doi: 10.1186/s13062-023-00449-8

Figure Lengend Snippet: GPR137 suppresses Hippo signaling by MST. A Protein levels of pMST1, MST1, pLATS1, LATS1, pLATS2 and LATS2 in AGS cells transfected with GPR137 (GPR137, +) or an empty vector (GPR137, -) for 24 h. One representative of 3 independent blots is shown. B Protein levels of pTAZ, TAZ, pYAP, YAP, CTGF and CYR61 in AGS cells transfected with GPR137 (GPR137, +) or an empty vector (GPR137, -) for 24 h. One representative of 3 independent blots is shown. C Co-immunoprecipitation of HA-tagged GPR137 and Flag-tagged MST1 in HEK293T cells. IP: Flag; WB: HA. N = 3. D Co-immunoprecipitation of endogenous GPR137 and MST1 in AGS cells. IP: MST1; WB: GPR137. IgG was used as a negative control. One representative of 3 independent blots is shown. E Co-immunoprecipitation of endogenous MST1 and LATS1 in the presence of GPR137 in AGS cells. IP: MST1; WB: LATS1. IgG was used as a negative control. One representative of 3 independent blots is shown. F Co-immunoprecipitation of endogenous MST1 and LATS1 in the presence of XMU-MP-1 in AGS cells. IP: MST1; WB: LATS1. IgG was used as a negative control. One representative of 3 independent blots is shown. G Protein expression examination of MST1 and MST2 in MST1/2 -knockout ( MST1/2 -KO) AGS cells or control AGS cells. One representative of 3 independent blots is shown. H Nucleo-cytoplasmic separation assays in MST1/2 -knockout AGS ( MST1/2 -KO) cells or control cells, and protein levels of YAP and TAZ were detected. Proteins in nucleus (left) and in cytoplasm (right). One representative of 3 independent blots is shown. I Nucleo-cytoplasmic separation assays in GPR137-transfected AGS (GPR137, +) cells or control cells (GPR137, -), and protein levels of YAP and TAZ were detected. Proteins in nucleus (upper) and in cytoplasm (lower). One representative of 3 independent blots is shown. J TEAD-luciferase assays in MST1/2 -knockout ( MST1/2 -KO) AGS cells or control AGS cells transfected with GPR137 or an empty vector (control). ** p < 0.01; error bar, SD. N = 3. K TEAD-luciferase assays in MST1/2 -knockout ( MST1/2 -KO) AGS cells or control AGS cells transfected with GPR137 shRNA or control scrambled shRNA. ** p < 0.01; error bar, SD. N = 3

Article Snippet: Phospho-MST1 (pMST1-Thr183, bs-4635R), MST1 (bs-3504R), MST2 (bs-4663R), phospho-LATS1 (pLATS1-Thr1079, bs-3245R), GPR137 (bs-16270R), PCNA (bs-2007R) antibodies were from Bioss (Beijing, China).

Techniques: Transfection, Plasmid Preparation, Immunoprecipitation, Negative Control, Expressing, Knock-Out, Control, Luciferase, shRNA

GPR137 loss declines AGS cell malignancy but potentiates Hippo activity . A Protein expression examination of GPR137 in GPR137 -knockout ( GPR137 -KO) AGS cells or control AGS cells. One representative of 3 independent blots is shown. B CCK-8 assays of GPR137-knockout AGS cells or control cells at different periods. ** p < 0.01; error bar, SD. N = 3. C Wound healing assays of GPR137 -knockout ( GPR137 -KO) AGS cells or control cells at 24 h. Bar = 100 μm. N = 3. D Statistical analysis of unoccupied area in ( C ). ** p < 0.01; error bar, SD. E Matrigel invasion assays of GPR137 -knockout ( GPR137 -KO) AGS cells or control cells at 24 h. Bar = 100 μm. N = 3. F Quantitative analysis of ( E ). ** p < 0.01; error bar, SD. G Colony formation assays of GPR137 -knockout ( GPR137 -KO) AGS cells or control cells. N = 3. H Quantitative analysis of (G). ** p < 0.01; error bar, SD. I Protein levels of pMST1 and MST1 in GPR137 -knockout ( GPR137 -KO) AGS cells or control cells. One representative of 3 independent blots is shown. J Protein levels of pTAZ, TAZ, pYAP and YAP in GPR137 -knockout ( GPR137 -KO) AGS cells or control cells. One representative of 3 independent blots is shown. K Nucleo-cytoplasmic separation assays in GPR137 -knockout AGS ( GPR137 -KO) cells or control cells, and protein levels of YAP and TAZ were detected. Proteins in nucleus (upper) and in cytoplasm (lower). One representative of 3 independent blots is shown. L , M qRT-PCR assays for mRNA levels of CTGF (L) and CYR61 (M) in GPR137 -knockout ( GPR137 -KO) AGS cells or control cells. ** p < 0.01; error bar, SD. N = 3. N Immunofluorescent staining for HA-derived signal in GPR137 -knockout ( GPR137 -KO) AGS cells or control cells transfected with HA-TAZ-S89A (upper) or HA-YAP-S127A (lower). Nuclei were stained with DAPI. Bar = 5 μm. N = 3

Journal: Biology Direct

Article Title: GPR137 inactivates Hippo signaling to promote gastric cancer cell malignancy

doi: 10.1186/s13062-023-00449-8

Figure Lengend Snippet: GPR137 loss declines AGS cell malignancy but potentiates Hippo activity . A Protein expression examination of GPR137 in GPR137 -knockout ( GPR137 -KO) AGS cells or control AGS cells. One representative of 3 independent blots is shown. B CCK-8 assays of GPR137-knockout AGS cells or control cells at different periods. ** p < 0.01; error bar, SD. N = 3. C Wound healing assays of GPR137 -knockout ( GPR137 -KO) AGS cells or control cells at 24 h. Bar = 100 μm. N = 3. D Statistical analysis of unoccupied area in ( C ). ** p < 0.01; error bar, SD. E Matrigel invasion assays of GPR137 -knockout ( GPR137 -KO) AGS cells or control cells at 24 h. Bar = 100 μm. N = 3. F Quantitative analysis of ( E ). ** p < 0.01; error bar, SD. G Colony formation assays of GPR137 -knockout ( GPR137 -KO) AGS cells or control cells. N = 3. H Quantitative analysis of (G). ** p < 0.01; error bar, SD. I Protein levels of pMST1 and MST1 in GPR137 -knockout ( GPR137 -KO) AGS cells or control cells. One representative of 3 independent blots is shown. J Protein levels of pTAZ, TAZ, pYAP and YAP in GPR137 -knockout ( GPR137 -KO) AGS cells or control cells. One representative of 3 independent blots is shown. K Nucleo-cytoplasmic separation assays in GPR137 -knockout AGS ( GPR137 -KO) cells or control cells, and protein levels of YAP and TAZ were detected. Proteins in nucleus (upper) and in cytoplasm (lower). One representative of 3 independent blots is shown. L , M qRT-PCR assays for mRNA levels of CTGF (L) and CYR61 (M) in GPR137 -knockout ( GPR137 -KO) AGS cells or control cells. ** p < 0.01; error bar, SD. N = 3. N Immunofluorescent staining for HA-derived signal in GPR137 -knockout ( GPR137 -KO) AGS cells or control cells transfected with HA-TAZ-S89A (upper) or HA-YAP-S127A (lower). Nuclei were stained with DAPI. Bar = 5 μm. N = 3

Article Snippet: Phospho-MST1 (pMST1-Thr183, bs-4635R), MST1 (bs-3504R), MST2 (bs-4663R), phospho-LATS1 (pLATS1-Thr1079, bs-3245R), GPR137 (bs-16270R), PCNA (bs-2007R) antibodies were from Bioss (Beijing, China).

Techniques: Activity Assay, Expressing, Knock-Out, Control, CCK-8 Assay, Quantitative RT-PCR, Staining, Derivative Assay, Transfection