Figure 1
Hypoxia induces secretion of high-mobility group box 1 (HMGB1) protein in normal human nasal epithelium (NHNE) cells. (a) NHNE cells were incubated under hypoxic conditions for 8 h and real-time PCR was performed to evaluate relative amount of HMGB1 mRNA. (b) NHNE cells were incubated under hypoxic conditions and western blot assay was performed to evaluate the relative amount of HMGB1 protein. (c) Relative band intensities of HMGB1 protein in whole-cell lysates were calculated and compared between control and hypoxic conditions (N=3). (d) NHNE cells were incubated under hypoxic conditions and then immunofluorescence assay was performed for HMGB1 (red) and with 4',6-diamidino-2-phenylindole (DAPI, blue) for DNA. Arrows indicate cytoplasmic HMGB1 protein. (e) Apical culture supernatants of NHNE cells were collected and concentrated. A western blot assay was performed to compare the HMGB1 protein level secreted into the extracellular space. (f) Relative band intensities of HMGB1 protein in culture supernatants were calculated and compared between control and hypoxic conditions (N=3). *P<0.05.
