Fig 1: Network analysis predicting NF-?B–deregulated PNs confirmed by histological and in vitro analyses.(A and B) Differentially expressed gene network plots (cellHarmony) showing the central hubs in the NMSC PN cluster (A) and SGCs (B), containing NF-?B transcription factors. Genes shown adjacent to red dots are upregulated, and those next to blue dots are downregulated in 7-month PNs versus 7-month control. (C and D) Immunostaining of tissue sections shows the NF-?B protein p65 (red; C) in mouse PN SCs (expressing EGFP; green) and activated, phosphorylated, p65 (red; D) in mouse PN SC nuclei (green). Yellow arrows indicate colocalization. Scale bars: 15 µm. (E) Normal human SCs (NHSCs) express less p60 and p50 than sphere-forming cells from human PNs (human sphere). In mouse, SCPs from embryonic DRGs contain less p60 and p50 compared with either SCP-like cells from PNs or mouse Nf1–/– SCPs. Lamin B1 was used as a loading control. See complete unedited blots in the supplemental material. (F and G) Numbers of mouse Nf1–/– embryonic SCP spheres (F) and human SCP-like cells (G) are slightly reduced by infection with a dominant negative NF-?B (I?B-SS). *P < 0.05 by Welch’s t test. (H) Western blot confirming downregulation of p65 by I?B-SS. (I) p65 immunoreactivity in many cells in human PN tissue sections (1:400). Inset: At 1:100, 38% of cells show immunoreactivity.
Supplier Page from MilliporeSigma for Anti-NFKB1 antibody produced in rabbit