Supplemental Experimental Procedures from The Beta Subunit of Hemoglobin (HBB2/HBB) Suppresses Neuroblastoma Growth and Metastasis

This file includes supplemental experimental procedures. It includes a description of animals, antibodies, dialysis of lung-derived factors, in-vitro viability assays, blocking the inhibitory effect of lung-derived factors, quantitative Real-Time PCR, fixation and sectioning of frozen tissues, immunofluorescence of frozen tissue sections, protein extraction from mouse organs, western blotting, enzyme-Linked Immunosorbent assay, Isolation of primary endothelial and epithelial cells from mouse lungs, Incubation of HPF and HPEC with MicroNB cells in Transwell system, apoptosis assay, cell cycle analysis, ERK1/2, p38 and TAK1 phosphorylation, Solid-phase synthesis of human HBB segments, Conjugation of Metox peptide to FITC, Confocal microscopy for Metox cell entry, Flow cytometry for Metox cell entry, orthotopic inoculation of tumor cells, monitoring tumor growth and metastasis, RNA preparation and Reverse Transcription Polymerase Chain Reaction, quantitative real-time PCR, intravenous and intranasal treatment of Metox, hemolytic assay, and nitric oxide assay.