Journal: Methods in molecular biology (Clifton, N.J.)

Article Title: Use of Inhibitors in the Study of MAP Kinases

doi: 10.1007/978-1-60761-795-2_6

Figure Lengend Snippet: List of antibodies for the phosphorylated and phosphorylation-independent (total) forms of MAP kinases and phosphorylated forms of MAP kinase substrate proteins

Article Snippet: , Stat-1 (S727) , CST (9177).

Techniques: Phospho-proteomics

Journal: American journal of physiology. Renal physiology

Article Title: The C-terminal tail of polycystin-1 regulates complement factor B expression by signal transducer and activator of transcription 1.

doi: 10.1152/ajprenal.00428.2015

Figure Lengend Snippet: Fig. 1. Polycystin-1 C-terminal tail (PC1- CTT), phosphorylated (p)-JAK2, p-STAT1, and complement factor B (CFB) expression in polycystic kidney disease (PKD). A: p-JAK2, total JAK2, p-STAT1, total STAT1, CFB, and PC1-CTT in control and human autosomal dominant (AD) PKD kidneys were analyzed by Western blotting and then quantified by densitometry. A representative Western blot is shown, and results represent an average of 3 independent experiments. NM, nonmutated. B: p-JAK2, total JAK2, p-STAT1, total STAT1, CFB, and PC1-CTT in / and Cy/ Han: SPRD rat kidneys were analyzed by Western blotting and then quantified by densitometry. Representative Western blot is shown, and results represent an average of 3 independent experiments. WT, wild-type. C: immunohisto- chemistry staining of p-JAK2, p-STAT1, CFB, and PC1-CTT in Han:SPRD rat kidneys is shown. One representative of 3 independent experiments is shown. Bars 40 m. NS, not significant. *P 0.05. **P 0.01. ***P 0.001.

Article Snippet: Total STAT1 (9172), p-JAK2 (3776), and p-STAT1 (7649) antibodies were from Cell Signaling Technology (Beverly, MA).

Techniques: Expressing, Control, Western Blot, Immunohistochemistry, Staining

Journal: American journal of physiology. Renal physiology

Article Title: The C-terminal tail of polycystin-1 regulates complement factor B expression by signal transducer and activator of transcription 1.

doi: 10.1152/ajprenal.00428.2015

Figure Lengend Snippet: Fig. 3. Effect of STAT1 on CFB expression. A: CFB expression in STAT1-overexpressed UCL93 renal epithelial cells 48 h after transfection. Blots were quantified by densitometry, and the average densitometry values from 3 independent experiments is shown. B: CFB expression in renal epithelial cells which were treated with the STAT1 inhibitor fludarabine for 48 h. Blots were quantified by densitometry, and the average densitometry values from 3 independent experiments are shown. One representative Western blot of 3 independent experiments is shown. *P 0.05. **P 0.01. ***P 0.001.

Article Snippet: Total STAT1 (9172), p-JAK2 (3776), and p-STAT1 (7649) antibodies were from Cell Signaling Technology (Beverly, MA).

Techniques: Expressing, Transfection, Western Blot

Journal: American journal of physiology. Renal physiology

Article Title: The C-terminal tail of polycystin-1 regulates complement factor B expression by signal transducer and activator of transcription 1.

doi: 10.1152/ajprenal.00428.2015

Figure Lengend Snippet: Fig. 4. A: representation of CFB promoter with putative STAT binding site (219/210). B: luciferase activity of CFB promoter (CFB-Pr) or mutated CFB promoter (CFB-Pr-Mut) was measured in control or STAT1-transfected UCL93 cells. C: luciferase activity of CFB-Pr, GAS promoter (GAS-Pr), or mutated GAS promoter (GAS-Pr-Mut) was measured in control or STAT1-transfected UCL93 cells. D: a chromatin immunoprecipitation (ChIP) assay was conducted of 293T cells transfected with control or STAT1. Two different regions (700/350, 400/100) were amplified by PCR after ChIP using an anti-STAT1 antibody. One representative of 3 independent experiments is shown. **P 0.01.

Article Snippet: Total STAT1 (9172), p-JAK2 (3776), and p-STAT1 (7649) antibodies were from Cell Signaling Technology (Beverly, MA).

Techniques: Binding Assay, Luciferase, Activity Assay, Control, Transfection, Chromatin Immunoprecipitation

Journal: American journal of physiology. Renal physiology

Article Title: The C-terminal tail of polycystin-1 regulates complement factor B expression by signal transducer and activator of transcription 1.

doi: 10.1152/ajprenal.00428.2015

Figure Lengend Snippet: Fig. 5. Effect of PC1-CTT on JAK2/STAT1 activation and CFB expression. A: JAK2/ STAT1 activation and CFB expression in UCL93 renal epithelial cell lines after 48-h PC1-CTT transfection. Blots were quantified by densitometry, and the average densitom- etry values from 3 independent experiments are shown. B: PC1-CTT was transfected in renal epithelial cells and followed by 24-h fludarabine treatment. CFB expression was an- alyzed by Western blotting and then quantified by densitometry. The average densitometry values from 3 independent experiments are shown. One representative Western blot of three independent experiments is shown. *P 0.05. **P 0.01. ***P 0.001.

Article Snippet: Total STAT1 (9172), p-JAK2 (3776), and p-STAT1 (7649) antibodies were from Cell Signaling Technology (Beverly, MA).

Techniques: Activation Assay, Expressing, Transfection, Western Blot

Journal: American journal of physiology. Renal physiology

Article Title: The C-terminal tail of polycystin-1 regulates complement factor B expression by signal transducer and activator of transcription 1.

doi: 10.1152/ajprenal.00428.2015

Figure Lengend Snippet: Fig. 6. A: luciferase activity of CFB-Pr or CFB-Pr-Mut was measured in control or PC1-CTT-transfected renal epithelial cells. B: cotransfection of renal epithelial cells with DN-STAT1 (DN1-STAT1 or DN2-STAT1) attenuated PC1-CTT-induced CFB-Pr activity. C: luciferase activity of CFB-Pr, GAS-Pr, or GAS-Pr-Mut was measured in control or PC1-CTT-transfected UCL93 cells. One representative of three independent experiments is shown. *P 0.05. **P 0.01.

Article Snippet: Total STAT1 (9172), p-JAK2 (3776), and p-STAT1 (7649) antibodies were from Cell Signaling Technology (Beverly, MA).

Techniques: Luciferase, Activity Assay, Control, Transfection, Cotransfection

Journal: American journal of physiology. Renal physiology

Article Title: The C-terminal tail of polycystin-1 regulates complement factor B expression by signal transducer and activator of transcription 1.

doi: 10.1152/ajprenal.00428.2015

Figure Lengend Snippet: Fig. 7. PC1-CTT promotes RAW264.7 macrophages toward M2 phenotype differentiation through STAT1 and CFB. A: bioactive human CFB was added to unconditioned media to treat macrophages. The expression of the macrophage M1 phenotype marker inducible nitric oxide synthase (iNOS) and M2 phenotype marker arginase-1 were examined by Western blotting. Blots were quantified by densitometry, and the average densitometry values from 3 independent experiments are shown. B: conditioned media were prepared from control (Control-CM) or PC1-CTT (PC1-CTT-CM)-transfected renal epithelial cells. Isotype IgG or CFB antibody was added to treat macrophages. C: conditioned media were generated from renal epithelial cells treated with fludarabine and used to treat macrophages for 24 h. D: PC1-CTT was transfected in renal epithelial cells and followed by fludarabine treatment, and conditioned media were collected to treat macrophages. One representative of 3 independent experiments is shown. *P 0.05. **P 0.01. ***P 0.001.

Article Snippet: Total STAT1 (9172), p-JAK2 (3776), and p-STAT1 (7649) antibodies were from Cell Signaling Technology (Beverly, MA).

Techniques: Expressing, Marker, Western Blot, Control, Transfection, Generated