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p 701 stat1  (Cell Signaling Technology Inc)
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Cell Signaling Technology Inc p 701 stat1
Figure 2. The role of KCa3.1 in M1 and M2 macrophage polarization in vitro. Macrophage colony-stimulating factor (M-CSF)–stimulated human macrophages were cotreated with vehicle or 100 nmol/L TRAM-34 during interferon (IFN)-γ and lipopolysaccharide LPS-induced M1 polarization and interleukin (IL)-4-induced M2 polarization. A, Flow cytometry analysis of cell surface markers for M1 (CD80 and CD197) and M2 (CD163 and CD206) macrophages during M1 and M2 polarization with or without TRAM-34. The flow cytometry results are expressed as MFI (n=3 in each group). B, Real-time polymerase chain reaction was performed to further evaluate the transcript expression of genes related to M1 and M2 macrophage polarization (n=5–6 in each group). C, Western blotting was conducted on cell lysates with antibodies for <t>P-STAT1,</t> P-STAT3, and P-STAT6; total STAT1, STAT3, and STAT6; and GAPDH. The relative densities for the phosphorylated STAT compared with total STAT are shown as histograms (n=4 in each group). Statistical significance values (P) are indi- cated. MFI indicates mean fluorescence intensity; and STAT, signal transducer and activator of transcription.
P 701 Stat1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Figure 2. The role of KCa3.1 in M1 and M2 macrophage polarization in vitro. Macrophage colony-stimulating factor (M-CSF)–stimulated human macrophages were cotreated with vehicle or 100 nmol/L TRAM-34 during interferon (IFN)-γ and lipopolysaccharide LPS-induced M1 polarization and interleukin (IL)-4-induced M2 polarization. A, Flow cytometry analysis of cell surface markers for M1 (CD80 and CD197) and M2 (CD163 and CD206) macrophages during M1 and M2 polarization with or without TRAM-34. The flow cytometry results are expressed as MFI (n=3 in each group). B, Real-time polymerase chain reaction was performed to further evaluate the transcript expression of genes related to M1 and M2 macrophage polarization (n=5–6 in each group). C, Western blotting was conducted on cell lysates with antibodies for P-STAT1, P-STAT3, and P-STAT6; total STAT1, STAT3, and STAT6; and GAPDH. The relative densities for the phosphorylated STAT compared with total STAT are shown as histograms (n=4 in each group). Statistical significance values (P) are indi- cated. MFI indicates mean fluorescence intensity; and STAT, signal transducer and activator of transcription.

Journal: Arteriosclerosis, thrombosis, and vascular biology

Article Title: Role of KCa3.1 Channels in Macrophage Polarization and Its Relevance in Atherosclerotic Plaque Instability.

doi: 10.1161/ATVBAHA.116.308461

Figure Lengend Snippet: Figure 2. The role of KCa3.1 in M1 and M2 macrophage polarization in vitro. Macrophage colony-stimulating factor (M-CSF)–stimulated human macrophages were cotreated with vehicle or 100 nmol/L TRAM-34 during interferon (IFN)-γ and lipopolysaccharide LPS-induced M1 polarization and interleukin (IL)-4-induced M2 polarization. A, Flow cytometry analysis of cell surface markers for M1 (CD80 and CD197) and M2 (CD163 and CD206) macrophages during M1 and M2 polarization with or without TRAM-34. The flow cytometry results are expressed as MFI (n=3 in each group). B, Real-time polymerase chain reaction was performed to further evaluate the transcript expression of genes related to M1 and M2 macrophage polarization (n=5–6 in each group). C, Western blotting was conducted on cell lysates with antibodies for P-STAT1, P-STAT3, and P-STAT6; total STAT1, STAT3, and STAT6; and GAPDH. The relative densities for the phosphorylated STAT compared with total STAT are shown as histograms (n=4 in each group). Statistical significance values (P) are indi- cated. MFI indicates mean fluorescence intensity; and STAT, signal transducer and activator of transcription.

Article Snippet: The antibodies used targeted KCa3.1 (Abcam, cat. no. ab83740), P-701-STAT1 (Cell Signaling, cat. no. 7649), STAT1 (Cell Signaling, cat. no. 9172), P-705-STAT3 (Cell Signaling, cat. no. 9145), STAT3 (Cell Signaling, cat. no. 4904), STAT 6 (Cell Signaling, cat. no.5397), P-641-STAT6 (Cell Signaling, cat. no. 9361) and GAPDH (Santa Cruz, cat. no. sc-365062).

Techniques: In Vitro, Flow Cytometry, Real-time Polymerase Chain Reaction, Expressing, Western Blot, Fluorescence