antibodies against phosphorylated and unphosphorylated human stat proteins  (Cell Signaling Technology Inc)

Journal: International journal of molecular sciences

Article Title: Cytokine Signaling in Pediatric Kidney Tumor Cell Lines WT-CLS1, WT-3ab and G-401.

doi: 10.3390/ijms25042281

Figure Lengend Snippet: Figure 1. The ARPE-19, WT-CLS1, WT-3ab and G-401 cells were treated with 10 ng/mL of cytokine for 20 min. The membranes were probed with phospho-specific antibodies recognizing only translocated STAT proteins (see Section 4); (A)—cells were stimulated with IFN-α and IFN-γ and probed with anti-phospho-STAT antibody; (B)—cells were stimulated with IFN- α and IL-6 and probed with anti- phospho-STAT3 antibody; (C)—cells were stimulated with IL-4 and probed with anti-phospho-STAT6 antibody. In addition, the membranes were probed with corresponding STAT antibodies, which recognize total STAT protein independent of phosphorylation (A–C—lower part). As a positive control, the lysates of ARPE-19 cells treated with appropriated cytokines were used. M—molecular weight marker.

Article Snippet: Primary rabbit polyclonal antibodies against phosphorylated and unphosphorylated human STAT proteins for Western blot experiments were from Cell Signaling (Danvers, MA, USA).

Techniques: Phospho-proteomics, Positive Control, Molecular Weight, Marker

Journal: International journal of molecular sciences

Article Title: Cytokine Signaling in Pediatric Kidney Tumor Cell Lines WT-CLS1, WT-3ab and G-401.

doi: 10.3390/ijms25042281

Figure Lengend Snippet: Figure 3. Flow cytometry analysis of WT-CLS1 cells stimulated with IFN-α, IFN-γ, IL-6 as well as with IL-4 and probed with anti-phospho-STAT antibodies. (A) (left part, blue histogram): Phospho-STAT1 staining. Fluorescence intensity of WT-CLS1 cells stained with isotype-matched control antibodies (negative control)—grey dotted histogram, unstimulated cells probed with anti-phospho-STAT1 antibody—grey histogram, WT-CLS1 cells stimulated with IFN-α (red histogram) and stimulated with IFN-γ (blue histogram). Middle part: After stimulation with IFN-α and IL-6 and analysis of phospho-STAT3. Grey dotted histogram—isotype-matched negative control. Unstimulated cells— grey histogram, cells stimulated with IFN-α (red histogram) and stimulated with IL-6 (green his- togram). Right part: After stimulation with IL-4 and analysis of phospho-STAT6. Grey dotted histogram—isotype-matched control. Unstimulated cells—grey histogram, stimulated with IL-6 (orange histogram). (B)—MHC class I and class II modulation. Left part: Flow cytometry analysis of WT-CLS1 cells stimulated with IFN-α and with IFN-γ for 48 h. The WT-CLS1 cells express high amounts of MHC class I (grey histogram), after stimulation with IFN-α the overexpression of MHC class I was observed (red histogram) and with IFN-γ (blue histogram). The grey dotted histograms represent isotype-matched negative controls. Right: MHC class II modulation. Flow cytometry analysis of WT-CLS1 cells stimulated with IFN-α and IFN-γ. The grey dotted histogram shows isotype-matched negative control. WT-CLS1 cells do not express MHC class II (grey histogram). Stimulation with IFN-α (red histogram) and IFN-γ (blue histogram) did not influence the MHC class II expression in WT-CLS1 cells. All four histograms overlap with each other.

Article Snippet: Primary rabbit polyclonal antibodies against phosphorylated and unphosphorylated human STAT proteins for Western blot experiments were from Cell Signaling (Danvers, MA, USA).

Techniques: Flow Cytometry, Staining, Fluorescence, Control, Negative Control, Over Expression, Expressing