human igg fc control  (R&D Systems)

Journal: bioRxiv

Article Title: NOTCH1 Acts as a Tumor Suppressor That Induces Early Differentiation in Head and Neck Cancer

doi: 10.1101/2025.04.25.650710

Figure Lengend Snippet: Ectopic expression of ICN1 mimics changes induced by JAG1 stimulation. A. Infection with ICN1 but not empty vector (MigR1) strongly inhibited protein expression of AXL and α-CATULIN. Widely used ICN1 cDNA product which begins several amino acids downstream of the native cleavage site is recognized by antibodies to the C-terminal region of NOTCH1 but not antibodies specific to the cleavage site. B. Expression of ICN1 in NOTCH1 mutant UMSCC22A produced the same morphological changes observed earlier upon expression of NFL1 and growth on JAG1. C. Expression of ICN1 in NOTCH1 WT 183 produced the same morphological changes observed earlier with growth on JAG1. D. Expression of ICN1 but not MigR1 control in NOTCH1 WT PJ34 produced the same morphological changes observed earlier with growth on JAG1. E. ICN1 expression triggered decreased protein expression of AXL, α-CATULIN, and ITGA3 in 183 cells.

Article Snippet: For NOTCH activation experiments, tissue culture wells were pre-coated overnight at room temperature (RT) with Protein G (Prospec, East Brunswick, NJ) at 50 ug/ml, washed twice in phosphate buffered saline (PBS), blocked in 1% BSA/PBS for 2 hours at RT, washed three times with PBS, coated with either human recombinant chimeric Jag1 fused to an FC fragment (R&D Systems, Minneapolis, MN) or control purified IgG FC protein (Jackson ImmunoResearch, West Grove, PA) at 2 ug/ml in 0.1% BSA/PBS for 3 hours RT, stored overnight at 4 0 C, and washed three times immediately before use.

Techniques: Expressing, Infection, Plasmid Preparation, Mutagenesis, Produced, Control

Journal: bioRxiv

Article Title: NOTCH1 Acts as a Tumor Suppressor That Induces Early Differentiation in Head and Neck Cancer

doi: 10.1101/2025.04.25.650710

Figure Lengend Snippet: Activation of NOTCH1 fails to increase surface CD133+ but increases expression of SOX2 in NOTCH1 mutant (UMSCC22A-iICN1), or NOTCH1 WT cell lines (FaDu-iICN1 and PJ34-iICN1). A. Cells were incubated for 48 h with or without DOX at 200 ng/ml (UMSCC22A), 300 ng/ml (FaDu), or 1000 ng/ml (PJ34) before staining with antibody to surface CD133 by flow cytometry. Fluorescent intensity histograms are shown for control biological replicates without DOX (red and light blue traces) or after DOX treatment (green and orange traces). B. Statistical comparison of CD133 mean fluorescence intensity (MFI). C. Western blot analysis of SOX2 protein expression in similarly treated cells.

Article Snippet: For NOTCH activation experiments, tissue culture wells were pre-coated overnight at room temperature (RT) with Protein G (Prospec, East Brunswick, NJ) at 50 ug/ml, washed twice in phosphate buffered saline (PBS), blocked in 1% BSA/PBS for 2 hours at RT, washed three times with PBS, coated with either human recombinant chimeric Jag1 fused to an FC fragment (R&D Systems, Minneapolis, MN) or control purified IgG FC protein (Jackson ImmunoResearch, West Grove, PA) at 2 ug/ml in 0.1% BSA/PBS for 3 hours RT, stored overnight at 4 0 C, and washed three times immediately before use.

Techniques: Activation Assay, Expressing, Mutagenesis, Incubation, Staining, Flow Cytometry, Control, Comparison, Fluorescence, Western Blot