Journal: Oncotarget

Article Title: NFATc3 plays an oncogenic role in oral/oropharyngeal squamous cell carcinomas by promoting cancer stemness via expression of OCT4

doi: 10.18632/oncotarget.26774

Figure Lengend Snippet: ( A ) Effect of NFATc3 on pluripotent transcription factors (NANOG, OCT4, KLF4, LIN28, and SOX2) expression was determined by qPCR. Their levels in NOKSI/NFATc3 were plotted as fold change against those in NOKSI/EV. * P < 0.001. ( B ) Effect of NFATc3 on OCT4 promoter activity was determined by luciferase promoter assay. Cells were transfected with pGL3-Basic (promoter-less) or pGL3 vectors containing the 1.5-kb upstream (-1545∼ -24) of Oct4. * P < 0.001. ( C ) Sequence analysis reveals a consensus NFAT binding site (5′-GGAAA-3′) at -1088 ∼ -1084 indicated by star (upper diagram). OSCC cells were lysed and performed a ChIP assay. The fragment (-1191∼ -1061) containing the NFAT binding site was enriched with NFATc3, and the fragment (-2930∼ -2783) was amplified as a control. * P < 0.01. ( D ) Correlation analysis of NFATc3 and OCT4 mRNA was determined based on their expression levels in 18 human SCC cell lines by qPCR.

Article Snippet: The mammalian pCMV6-Entry Vector encoding OCT4 was purchased from OriGene (RC211998).

Techniques: Expressing, Activity Assay, Luciferase, Promoter Assay, Transfection, Sequencing, Binding Assay, Amplification

Journal: Oncotarget

Article Title: NFATc3 plays an oncogenic role in oral/oropharyngeal squamous cell carcinomas by promoting cancer stemness via expression of OCT4

doi: 10.18632/oncotarget.26774

Figure Lengend Snippet: ( A ) The effect of OCT4 knockdown on self-renewal capacity of NOKSI/NFATc3 was determined by tumor sphere formation assay. OCT4 was knocked down in NOKSI/NFATc3 using siRNA against OCT4 (Oct4i). The cells transfected with control siRNA (CTLi) were included for comparison. * P < 0.05. ( B ) The effect of OCT4 knockdown on migration ability in NOKSI/NFATc3 was determined by transwell migration assay. ** P < 0.01. ( C ) The effect of OCT4 knockdown on self-renewal capacity of SCC4 was determined by tumor sphere formation assay. ( D ) The effect of OCT4 knockdown on self-renewal capacity of SCC4 was determined by transwell migration assay. ( E ) OCT4 expression was forced in non-tumorigenic immortalized oral epithelial cells, NOKSI, by vector expressing recombinant Myc-DDK-tagged OCT4, and its ectopic expression was confirmed by Western blot analysis using anti-DDK antibody. ( F ) Effect of ectopic OCT4 expression on self-renewal capacity of NOKSI was determined by tumor sphere formation assay. Representative images of tumor spheres formed by NOKSI/EV and NOKSI/Oct4 are shown on the right. ( G ) Effect of ectopic OCT4 expression on migration ability of NOKSI was determined by transwell migration assay. ( H ) Effect of ectopic OCT4 expression on the expression of NFAT isoforms (NFATc1-c4) in NOKSI was determined by qPCR. Their levels in NOKSI/Oct4 were plotted as fold induction against those in NOKSI/EV.

Article Snippet: The mammalian pCMV6-Entry Vector encoding OCT4 was purchased from OriGene (RC211998).

Techniques: Tube Formation Assay, Transfection, Migration, Transwell Migration Assay, Expressing, Plasmid Preparation, Recombinant, Western Blot

Journal: PLoS ONE

Article Title: Identification of Rabbit Annulus Fibrosus-Derived Stem Cells

doi: 10.1371/journal.pone.0108239

Figure Lengend Snippet: The AF-derived cells were positive for Oct-4 ( A–C ), nucleostemin ( D–F ) and SSEA-4 ( G–I ). Scale bars, 200 µm.

Article Snippet: They were then blocked with 4% BSA for 30 min before being incubated with mouse anti-human Oct-4 antibody (1∶500, Millipore, Cat.# MAB4401), goat anti-human nucleostemin antibody (1∶250, Neuromics, Cat.# GT15050), and mouse anti-human SSEA-4 antibody (1∶200, Invitrogen, Cat.# 41-4000), for staining of Oct-4, nucleostemin, and SSEA-4, respectively.

Techniques: Derivative Assay