plasmid encoding human oct4 gene  (Addgene inc)

Journal: Cancer Management and Research

Article Title: OCT4 accelerates tumorigenesis through activating JAK/STAT signaling in ovarian cancer side population cells

doi: 10.2147/CMAR.S180418

Figure Lengend Snippet: OCT4 was overexpressed in the SP of ovarian cancer cells. Notes: ( A–C ) Western blotting and RT-PCR were carried out to analyze the protein and mRNA expressions of OCT4 in the SP and NSP population of SKOV3 and A2780 cells. ** P < 0.01; *** P <0.001. Abbreviations: NSP, non-SP; SP, side population.

Article Snippet: For knockdown of the expression of OCT4 stably, the shR-NAs targeting human OCT4 (No. TR310267) gene was purchased from OriGene (Rockville, MD, USA).

Techniques: Western Blot, Reverse Transcription Polymerase Chain Reaction

Journal: Cancer Management and Research

Article Title: OCT4 accelerates tumorigenesis through activating JAK/STAT signaling in ovarian cancer side population cells

doi: 10.2147/CMAR.S180418

Figure Lengend Snippet: Downregulation of OCT4 reduced cell drug resistance and inhibited cell proliferation and tumorigenesis in the SP of ovarian cancer cells. Notes: ( A , B ) Western blotting analysis of the knockdown efficiency of OCT4 after 48 hours of the cells were transfected with sh-OCT4. ( C , D ) Different concentrations of DDP were added in the SP of SKOV3 and A2780 cells after 48 hours of the cells were transfected with sh-OCT4, then CCK-8 assay was performed to assess cell viability. ( E , F ) CCK-8 analysis of cell viability after 48 hours of cell treatments. ( G , H ) Flow cytometry analysis of cell cycle after 48 hours of cell treatments. ( I , J ) In vivo xenograft model was carried out to analyze the effect of sh-OCT4 on tumorigenesis in the SP cells. The data presented are the mean ± standard error and represent three independent experiments (* P <0.05; ** P <0.01). Effects of downregulation of OCT4 on cell viability and cycle in the SP population of SKOV3 cells. Abbreviations: CCK-8, cell counting kit-8; SP, side population.

Article Snippet: For knockdown of the expression of OCT4 stably, the shR-NAs targeting human OCT4 (No. TR310267) gene was purchased from OriGene (Rockville, MD, USA).

Techniques: Western Blot, Transfection, CCK-8 Assay, Flow Cytometry, In Vivo, Cell Counting

Journal: Cancer Management and Research

Article Title: OCT4 accelerates tumorigenesis through activating JAK/STAT signaling in ovarian cancer side population cells

doi: 10.2147/CMAR.S180418

Figure Lengend Snippet: Effects of OCT4 overexpression on cell functions and drug resistance in the NSP of SKOV3 and A2780 cells. Notes: ( A , B ) Western blotting was carried out to analyze the protein expressions of OCT4 after 48 hours of the SP cells were treated with Lentiv-OCT4 or Lentiv-NC, respectively. ( C , D ) Different concentrations of DDP were added in the NSP of SKOV3 and A2780 cells after 48 hours of the cells were treated with Lentiv-OCT4 or Lentiv-NC, and then CCK-8 assay was performed to assess cell viability. ( E , F ) CCK-8 analysis of cell proliferation after 48 hours of the NSP cells was infected with Lentiv-OCT4 or Lentiv-NC. ( G , H ) Flow cytometry was used to assess cell cycle after 48 hours of the NSP cells were infected with Lentiv-OCT4 or Lentiv-NC. The data presented are the mean ± standard error and represent three independent experiments (* P <0.05; ** P <0.01). Abbreviations: CCK-8, cell counting kit-8; NSP, non-SP; SP, side population.

Article Snippet: For knockdown of the expression of OCT4 stably, the shR-NAs targeting human OCT4 (No. TR310267) gene was purchased from OriGene (Rockville, MD, USA).

Techniques: Over Expression, Western Blot, CCK-8 Assay, Infection, Flow Cytometry, Cell Counting

Journal: Cancer Management and Research

Article Title: OCT4 accelerates tumorigenesis through activating JAK/STAT signaling in ovarian cancer side population cells

doi: 10.2147/CMAR.S180418

Figure Lengend Snippet: Evaluation of the effect of OCT4 on JAK/STAT signaling pathway activation in the NSP of SKOV3 and A2780 cells. Notes: ( A , B ) Western blotting analysis of the protein expressions of phosphorylated JAK1 (p-JAK1), p-STAT6, p-AKT, and p-NF-κB after 48 hours of the NSP cells was infected with Lentiv-OCT4 or Lentiv-NC. ( C , D ) Western blotting analysis of the protein expressions of p-JAK1, p-JAK2, p-JAK3, and p-Tyk2 after 48 hours of the NSP cells was infected with Lentiv-OCT4 or Lentiv-NC. ( E , F ) Western blotting analysis of the protein expressions of p-STAT1, p-STAT2, p-STAT3, p-STAT4, p-STAT5, and p-STAT6 after 48 hours of the NSP cells was treated with Lentiv-OCT4 or Lentiv-NC. ( G , H ) Immunofluorescence staining was used to determine the subcellular location of STAT6 after 48 hours of the SP SKOV3 cells were transfected Lentiv-OCT4. ( I , J ) Western blotting analysis of the expression of Cyclin D1, c-Myc, and Bcl-2 after 48 hours of the NSP cells was treated with Lentiv-OCT4 or Lentiv-NC. The data presented are the mean ± standard error and represent three independent experiments (* , # P <0.05). Abbreviation: NSP, nonside population.

Article Snippet: For knockdown of the expression of OCT4 stably, the shR-NAs targeting human OCT4 (No. TR310267) gene was purchased from OriGene (Rockville, MD, USA).

Techniques: Activation Assay, Western Blot, Infection, Immunofluorescence, Staining, Transfection, Expressing

Journal: Cancer Management and Research

Article Title: OCT4 accelerates tumorigenesis through activating JAK/STAT signaling in ovarian cancer side population cells

doi: 10.2147/CMAR.S180418

Figure Lengend Snippet: Assessment of the effects of OCT4/JAK/STAT on cell functions in the NSP of SKOV3 and A2780 cells. Notes: ( A, B ) Western blotting analysis of the protein levels of JAK1 and caspase-3 in different treated NSP cells: Lentiv-NC (48 hours), Lentiv-OCT4 (48 hours), and peficitinib (4.8 nM, 2 hours) + Lentiv-OCT4 (48 hours). ( C , D ) Flow cytometry was performed to assess the apoptosis of the NSP cells with different treatments: Lentiv-NC (48 hours), Lentiv-OCT4 (48 hours), and peficitinib (4.8 nM, 2 hours) + Lentiv-OCT4 (48 hours). ( E , F ) CCK-8 and clone formation assays were performed to assess cell viability in the SP cells with different treatments: Lentiv-NC (48 hours), Lentiv-OCT4 (48 hours), and peficitinib (4.8 nM, 2 hours) + Lentiv-OCT4 (48 hours). ( G , H ). Transwell assay was used to evaluate the invasion of the NSP cells with different treatments: Lentiv-NC (48 hours), Lentiv-OCT4 (48 hours), and peficitinib (4.8 nM, 2 hours) + Lentiv-OCT4 (48 hours). The data presented are the mean ± standard error and represent three independent experiments (* , # P <0.05). Abbreviations: CCK-8, cell counting kit-8; NSP, nonside population.

Article Snippet: For knockdown of the expression of OCT4 stably, the shR-NAs targeting human OCT4 (No. TR310267) gene was purchased from OriGene (Rockville, MD, USA).

Techniques: Western Blot, Flow Cytometry, CCK-8 Assay, Transwell Assay, Cell Counting

Journal: Cancer Management and Research

Article Title: OCT4 accelerates tumorigenesis through activating JAK/STAT signaling in ovarian cancer side population cells

doi: 10.2147/CMAR.S180418

Figure Lengend Snippet: Detection of the effect of OCT4/JAK/STAT on the tumorigenesis of the NSP of SKOV3 and A2780 cells. Notes: In vivo xenograft model analysis of the effect of OCT4/JAK/STAT on tumorigenesis. The data presented are the mean ± standard error and represent three independent experiments (* , # P <0.05). Abbreviation: NSP, nonside population.

Article Snippet: For knockdown of the expression of OCT4 stably, the shR-NAs targeting human OCT4 (No. TR310267) gene was purchased from OriGene (Rockville, MD, USA).

Techniques: In Vivo

Journal: Cancer Management and Research

Article Title: OCT4 accelerates tumorigenesis through activating JAK/STAT signaling in ovarian cancer side population cells

doi: 10.2147/CMAR.S180418

Figure Lengend Snippet: Graphical abstract of this study. Notes: OCT4 activates JAK/STAT signaling, then promotes the expression of Cyclin D1, c-Myc, and Bcl-2, accelerating the tumorigenesis of NSP cells in ovarian cancer. Abbreviation: NSP, nonside population.

Article Snippet: For knockdown of the expression of OCT4 stably, the shR-NAs targeting human OCT4 (No. TR310267) gene was purchased from OriGene (Rockville, MD, USA).

Techniques: Expressing

Journal: Cancers

Article Title: An FDA-Approved Antifungal, Ketoconazole, and Its Novel Derivative Suppress tGLI1-Mediated Breast Cancer Brain Metastasis by Inhibiting the DNA-Binding Activity of Brain Metastasis-Promoting Transcription Factor tGLI1

doi: 10.3390/cancers14174256

Figure Lengend Snippet: KCZ and the novel derivative KCZ-7 inhibit tGLI1 transcriptional activity leading to downregulation of validated tGLI1-mediated stemness genes Nanog and OCT4 . ( a ) Representative Western blots of GLI1 and tGLI1 expression in isogenic SKBRM cell lines following 24 h treatment with vehicle, 1 μM KCZ, or 1 μM KCZ-7. The same membrane was probed to assess the loading control. ( b ) Western blots of recombinant GLI1 and N-tGLI1 (left). A tGLI1-selective Ab was used to detect tGLI1. Binding of recombinant GLI1 and N-tGLI1 to a dsDNA oligonucleotide containing the consensus GLI1/tGLI1-binding site (right). STAT3 was used as a negative control. ( c ) The DNA-binding ability of recombinant N-tGLI1, but not GLI1, is disrupted by KCZ or KCZ-7 treatment. ( d ) Relative binding of GLI1 or tGLI1 to the GLI1-binding sites in SKBRM cells, as determined by chromatin immunoprecipitation; qPCR was performed using primers spanning the GLI1 binding site. ( e , f ) Inhibition of GLI1- and tGLI1-mediated promoter transactivation by KCZ ( e ) and KCZ-7 ( f ). SKBR3 cells were transiently transfected with 8 × 3′GLI1 luciferase reporter and vector, GLI1, or tGLI1 plasmids, then treated with increasing doses of KCZ ( e ) or KCZ-7 ( f ) for 48 h and stimulated with SHH ligand (100 ng/mL) for 4 h. Right: Relative luciferase activity normalized to vehicle treatment. ( g , h ) Selective reduction of tGLI1-mediated stemness genes Nanog ( g ) and OCT4 ( h ) mRNA as assessed by RT-qPCR in isogenic SKBRM cell lines treated with vehicle, 1 μM KCZ, or 1 μM KCZ-7 for 24 h. ( i ) Nanog and OCT4 protein expression following treatment with vehicle, 1 μM KCZ, or 1 μM KCZ-7 in isogenic SKBRM cell lines. The same membrane was probed to assess the loading control. ( j , k ) Overexpression of Nanog ( j ) or OCT4 ( k ) rescues SKBRM-tGLI1 mammospheres from KCZ and KCZ-7 treatment. Scale bars represent 200 μm. N-tGLI1, N-terminal tGLI1; *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; two-way ANOVA with post hoc Dunnett’s ( d – f ) or Bonferroni’s ( g , h , j , k ) multiple comparison test was used to calculate p -values. The uncropped blots are shown in page 2 of .

Article Snippet: The Nanog (HG13138-UT) and OCT4 (HG13137-UT) overexpression plasmids were purchased from Sino Biological (Beijing, China).

Techniques: Activity Assay, Western Blot, Expressing, Recombinant, Binding Assay, Negative Control, Chromatin Immunoprecipitation, Inhibition, Transfection, Luciferase, Plasmid Preparation, Quantitative RT-PCR, Over Expression