Figure 1.
Primary culture of human nasal epithelial cells (HNEC).
Bright-field images 5 daysaftercell sampling (left),and 9 days after sampling with early epithelioid morphology (right).
Figure 2.
Expression of GFP in HNECs following various multiplicities of infection (MOI).
Induction of GFP protein with MOI = 1 (top), MOI = 2, (middle), and MOI = 3 (bottom). GFP expression was observed when MOI = 1 and 2, but the expression was stronger when MOI = 3.
Figure 3.
Schema of generation of iPS from HNECs.
Figure 4.
Morphologies of HNEC-derived iPS.
A. Colonies generated from HNECs show a round shape with large nucleoli and scant cytoplasm, similar to the morphologyof human ES cells. B. Crystal violet staining of growing cells. Efficiency of iPS cell generation is 0.75% at MOI 3 (left), and 0.1% at MOI 4 (right).
Figure 5.
Gene/Protein expressions of HNEC-derived iPS.
A. RT-PCR analyses for the hESC marker genes NANOG, OCT3/4, SOX2, KLF4, c-MYC, GDF3, REX31, DPPA2, and DPPA4 and the transgenes OCT3/4, SOX2, KLF4, and c-MYC. B. Immunofluorescence staining for pluripotency and surface markers (OCT4, SOX2, NANOG,SSEA4,Tra-1-60, and Tra-1-81). Scale bars indicate 500 µm. Red: Alexa 568, Green: Alexa488.
Figure 6.
Methylation analysis of the promoter region.
Bisulfite sequencing analysis of the NANOG and OCT3/4 promoter regions in HNECs, hESCs, and HNEC-derived iPS cells B2, B3, 2B1, 3A2, and 4B1 are shown. Open circles indicate unmethylated CpGs, while closed circles indicate methylated CpGs.
Figure 7.
Global gene expression analysis.
Gene expression patterns were compared between HNEC-derived iPS cells (iPS-B3) and HNECs (left, top), between HNEC-derived iPS cells (iPS-2B1) and HNECs (left, bottom), between HNEC-derived iPS cells (iPS-B3) and human ES cells (hESC, right, top), and between HNEC-derived iPS cells (iPS-2B1) and hESCs (right, bottom). Blue dotted lines indicate 2-fold changes. r represents the correlation coefficient.