Journal: International Journal of Nanomedicine

Article Title: Dual functions of silver nanoparticles in F9 teratocarcinoma stem cells, a suitable model for evaluating cytotoxicity- and differentiation-mediated cancer therapy

doi: 10.2147/IJN.S145147

Figure Lengend Snippet: Cytotoxicity assessment of AgNPs in F9 cells. Notes: ( A ) Viability of F9 cells was determined 24 h after exposure to different concentrations of AgNPs, using the WST-8 assay. The results are expressed as the mean ± standard deviation of three independent experiments. A significant difference was observed for AgNPs concentration above 6.125 μg/mL. ( B ) LDH activity was measured at 490 nm using the LDH cytotoxicity kit. The results are expressed as the mean ± standard deviation of three independent experiments. The viability of treated and untreated cells was compared using Student’s t -test ( P <0.05). There was a significant difference observed in the LDH activity of AgNPs-treated cells compared to that of the untreated cells using Student’s t -test (* P <0.5). Abbreviations: AgNPs, silver nanoparticles; LDH, lactate dehydrogenase.

Article Snippet: F9 mouse embryonic carcinoma cells were purchased from the Korean Cell Line Bank (KCLB) and maintained in DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution.

Techniques: Standard Deviation, Concentration Assay, Activity Assay

Journal: International Journal of Nanomedicine

Article Title: Dual functions of silver nanoparticles in F9 teratocarcinoma stem cells, a suitable model for evaluating cytotoxicity- and differentiation-mediated cancer therapy

doi: 10.2147/IJN.S145147

Figure Lengend Snippet: Effect of AgNPs on ROS generation. Notes: ( A ) F9 cells were treated with or without 12.5 μg/mL of AgNPs or 6.125 μg/mL of AgNO 3 for 24 h, and ROS generation was measured using flow cytometry. ( B ) Treated cells were measured for DCFH-DA-FITC by fluorescence microscopy analysis. Scale bar =200 μm. ( C ) The bar graph indicates the ratio between control and treated samples. The results are expressed as the mean ± standard deviation of three independent experiments. There was a significant difference observed in the ROS generation in the AgNPs- or AgNO 3 -treated cells compared to that of the untreated cells using Student’s t -test (* P <0.5, *** P <0.01). Abbreviations: AgNPs, silver nanoparticles; ROS, reactive oxygen species; DCFH-DA, 2′,7′-dichlorodihydrofluorescein diacetate; BF, bright field; FITC, fluorescein isothiocyanate.

Article Snippet: F9 mouse embryonic carcinoma cells were purchased from the Korean Cell Line Bank (KCLB) and maintained in DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution.

Techniques: Flow Cytometry, Fluorescence, Microscopy, Control, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Dual functions of silver nanoparticles in F9 teratocarcinoma stem cells, a suitable model for evaluating cytotoxicity- and differentiation-mediated cancer therapy

doi: 10.2147/IJN.S145147

Figure Lengend Snippet: AgNPs-induced loss of mitochondrial membrane potential. Notes: ( A ) F9 cells were treated with or without 12.5 μg/mL of AgNPs or 6.125 μM of AgNO 3 for 24 h. JC-1 monomer (green) and aggregate (red) formation was measured using flow cytometry. ( B ) Representative fluorescence images of JC-1 monomer/aggregate formation. ( C ) The bar graph indicates the JC-1 monomer:JC-1 aggregate formation ratio. The results are expressed as the mean ± standard deviation of three independent experiments. There was a significant difference observed in the ratio for AgNPs- or AgNO 3 -treated cells compared to that for the untreated cells by the Student’s t -test (* P <0.5, and *** P <0.01). Abbreviation: AgNPs, silver nanoparticles.

Article Snippet: F9 mouse embryonic carcinoma cells were purchased from the Korean Cell Line Bank (KCLB) and maintained in DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution.

Techniques: Membrane, Flow Cytometry, Fluorescence, Standard Deviation

Journal: International Journal of Nanomedicine

Article Title: Dual functions of silver nanoparticles in F9 teratocarcinoma stem cells, a suitable model for evaluating cytotoxicity- and differentiation-mediated cancer therapy

doi: 10.2147/IJN.S145147

Figure Lengend Snippet: AgNPs-induced apoptosis in F9 cells. Notes: ( A ) F9 cells were treated with or without 12.5 μg/mL of AgNPs or 6.125 μM of AgNO 3 for 24 h. Then, the apoptosis of cells was assessed using the TUNEL assay; the nuclei were counterstained with DAPI. Representative images show apoptotic (fragmented) DNA (red staining) and the corresponding cell nuclei (blue staining). ( B ) Apoptotic efficiency was measured using flow cytometry. Abbreviations: AgNPs, silver nanoparticles; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling.

Article Snippet: F9 mouse embryonic carcinoma cells were purchased from the Korean Cell Line Bank (KCLB) and maintained in DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution.

Techniques: TUNEL Assay, Staining, Flow Cytometry

Journal: International Journal of Nanomedicine

Article Title: Dual functions of silver nanoparticles in F9 teratocarcinoma stem cells, a suitable model for evaluating cytotoxicity- and differentiation-mediated cancer therapy

doi: 10.2147/IJN.S145147

Figure Lengend Snippet: Effect of AgNPs on pro- and antiapoptotic gene expression. Notes: ( A ) F9 cells were treated with or without 12.5 μg/mL of AgNPs or 6.125 μM of AgNO 3 for 24 h, and the relative mRNA expression was analyzed by RT-qPCR. ( B ) Protein expression was analyzed using western blot. ( C ) The bar graph indicates the signal intensity ratio between control and treated groups. The results are expressed as the mean ± standard deviation of three separate experiments. The treated groups showed statistically significant differences from the control group determined using Student’s t -test (* P <0.05 and ** P <0.1). Abbreviations: AgNPs, silver nanoparticles; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Article Snippet: F9 mouse embryonic carcinoma cells were purchased from the Korean Cell Line Bank (KCLB) and maintained in DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution.

Techniques: Gene Expression, Expressing, Quantitative RT-PCR, Western Blot, Control, Standard Deviation, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: International Journal of Nanomedicine

Article Title: Dual functions of silver nanoparticles in F9 teratocarcinoma stem cells, a suitable model for evaluating cytotoxicity- and differentiation-mediated cancer therapy

doi: 10.2147/IJN.S145147

Figure Lengend Snippet: Effect of AgNPs on expression of differentiation markers. Notes: ( A ) F9 cells were treated with or without RA (1 μM) for 72 h or AgNPs (6.125 or 12.5 μg/mL) for 24 h, and differentiation was observed by phase contrast microscopy. ( B ) Real-time quantitative PCR was performed to analyze the expression of various neuronal differentiation markers. The results are expressed as the mean ± standard deviation of three separate experiments. The treated groups showed statistically significant differences from the control group determined using Student’s t -test (*** P <0.001). Abbreviations: AgNPs, silver nanoparticles; RA, retinoic acid; PCR, polymerase chain reaction.

Article Snippet: F9 mouse embryonic carcinoma cells were purchased from the Korean Cell Line Bank (KCLB) and maintained in DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution.

Techniques: Expressing, Microscopy, Real-time Polymerase Chain Reaction, Standard Deviation, Control, Polymerase Chain Reaction

Journal: International Journal of Nanomedicine

Article Title: Dual functions of silver nanoparticles in F9 teratocarcinoma stem cells, a suitable model for evaluating cytotoxicity- and differentiation-mediated cancer therapy

doi: 10.2147/IJN.S145147

Figure Lengend Snippet: Effect of AgNPs on expression of stem cell markers. Notes: ( A ) F9 cells were treated with or without RA (1 μM) for 72 h or AgNPs (3.0625 to 50 μg/mL) for 24 h. Flow cytometry was performed to analyze the expression of the Nanog stem cell marker. ( B ) Real-time quantitative PCR was performed to analyze the expression of various other stem cell markers. The results are expressed as the mean ± standard deviation of three separate experiments. The treated groups showed statistically significant differences from the control group determined using the Student’s t -test (* P <0.05, ** P <0.01, and *** P <0.001). Abbreviations: AgNPs, silver nanoparticles; RA, retinoic acid; PCR, polymerase chain reaction; NS, not significant.

Article Snippet: F9 mouse embryonic carcinoma cells were purchased from the Korean Cell Line Bank (KCLB) and maintained in DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution.

Techniques: Expressing, Flow Cytometry, Marker, Real-time Polymerase Chain Reaction, Standard Deviation, Control, Polymerase Chain Reaction

Journal: International Journal of Nanomedicine

Article Title: Dual functions of silver nanoparticles in F9 teratocarcinoma stem cells, a suitable model for evaluating cytotoxicity- and differentiation-mediated cancer therapy

doi: 10.2147/IJN.S145147

Figure Lengend Snippet: Cisplatin reduces viability of F9 cells that underwent AgNPs-induced differentiation. Notes: F9 cells differentiated using RA (1 μM) for 72 h or AgNPs (6.125 or 12.5 μg/mL) for 24 h were treated with or without cisplatin (1 μM) for another 24 h, and then cell viability was measured. The results are expressed as the mean ± standard deviation of three separate experiments. The treated groups showed statistically significant differences from the control group determined using Student’s t -test (* P <0.05 and ** P <0.01). Abbreviations: AgNPs, silver nanoparticles; RA, retinoic acid.

Article Snippet: F9 mouse embryonic carcinoma cells were purchased from the Korean Cell Line Bank (KCLB) and maintained in DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution.

Techniques: Standard Deviation, Control

Journal: International Journal of Nanomedicine

Article Title: Dual functions of silver nanoparticles in F9 teratocarcinoma stem cells, a suitable model for evaluating cytotoxicity- and differentiation-mediated cancer therapy

doi: 10.2147/IJN.S145147

Figure Lengend Snippet: Role of cisplatin in AgNPs-induced expression of differentiation markers. Notes: ( A ) F9 cells differentiated using RA (1 μM) for 72 h or AgNPs (6.125 or 12.5 μg/mL) for 24 h were treated with or without cisplatin (1 μM) for another 24 h, and then the expression of various proteins involved in differentiation was analyzed using western blot. Results were normalized to β-actin. The results are expressed as the mean ± standard deviation of three separate experiments. ( B ) The bar graph indicates the signal intensity ratio between control and treated groups. The treated groups showed statistically significant differences from the control group determined using Student’s t-test (* P <0.05). L, represents low concentration of AgNPs, H, represents high concentration of AgNPs. Abbreviations: AgNPs, silver nanoparticles; RA, retinoic acid.

Article Snippet: F9 mouse embryonic carcinoma cells were purchased from the Korean Cell Line Bank (KCLB) and maintained in DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution.

Techniques: Expressing, Western Blot, Standard Deviation, Control, Concentration Assay

Journal: International Journal of Nanomedicine

Article Title: Dual functions of silver nanoparticles in F9 teratocarcinoma stem cells, a suitable model for evaluating cytotoxicity- and differentiation-mediated cancer therapy

doi: 10.2147/IJN.S145147

Figure Lengend Snippet: Role of cisplatin in AgNPs-modulated expression of proteins of various proliferation markers. Notes: ( A ) F9 cells differentiated using RA (1 μM) for 72 h or AgNPs (6.125 or 12.5 μg/mL) for 24 h were treated with or without cisplatin (1 μM) for 24 h, and then the expression of various proteins involved in proliferation was analyzed using western blot. Results were normalized to β-actin. The results are expressed as the mean ± standard deviation of three separate experiments. ( B ) The bar graph indicates the signal intensity ratio between control and treated groups. The treated groups showed statistically significant differences from the control group determined using Student’s t-test (* P <0.05). L, represents low concentration of AgNPs, H, represents high concentration of AgNPs.

Article Snippet: F9 mouse embryonic carcinoma cells were purchased from the Korean Cell Line Bank (KCLB) and maintained in DMEM supplemented with 10% FBS and 1% antibiotic–antimycotic solution.

Techniques: Expressing, Western Blot, Standard Deviation, Control, Concentration Assay

Journal: Cell Journal (Yakhteh)

Article Title: Differential Expression of OCT4 Pseudogenes in Pluripotent and Tumor Cell Lines

doi:

Figure Lengend Snippet: A. Schematic representation of OCT4 pseudogenes. OCT4-pg1, OCT4-pg3 and OCT4-pg4 have highly similar nucleotide sequences to that of the OCT4A transcript. OCT4-pg5 transcript lacks exon1, and OCT4-pg7 lacks exon1, exon4, and part of exon2. OCT4-pg2 has a part of exon5, and OCT4-pg6 has all five exons, incompletely. Rough lines in OCT4-pg2 and OCT4-pg6 are remained sequences which are derived from OCT4 introns and B. RT-PCR analysis of OCT4 -pseudogenes in different human pluripotent and cancer cell lines by specific primer sets. GAPDH was used as an internal control. RT-PCR; Reverse transcriptase-polymerase chain reaction.

Article Snippet: In this experimental study, 17 human cell lines were mainly provided by Pasteur Institute of Iran and Research Institute of Avicenna, included two human embryonic carcinoma cell lines, one normal fibroblast cell line (HS-5) and HEK293 (embryonic kidney) and 13 human tumor cell lines, namely U87MG and A172 (glioblastoma), Daoy (medulloblastoma), 1321N1 (brain astrocytoma), Jurkat (T-Cell lymphoma), Y79 (retinoblastoma), PC3 (prostate adenocarcinoma), Raji (Burkit’s lymphoma), Ovcar3 (ovary adenocarcinoma), HepG2 (hepatocellular carcinoma), MCF7 (breast adenocarcinoma), 5637 (urinary bladder carcinoma), HeLa (cervix adenocarcinoma), and NT2 and NCCIT (pluripotent embryonic carcinoma).

Techniques: Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Control, Reverse Transcription, Polymerase Chain Reaction

Journal: Cell Journal (Yakhteh)

Article Title: Differential Expression of OCT4 Pseudogenes in Pluripotent and Tumor Cell Lines

doi:

Figure Lengend Snippet: A. A schematic view of OCT4A protein structure, along with predicted protein structures of OCT4-Pg1, OCT4-Pg3 , and OCT4-Pg4 . The putative OCT4-Pg1 protein is similar to OCT4A, containing intact NTD, POU domain and CTD. OCT4-Pg3 can potentially produce a truncated protein with NTD and a part of the POUs domain. The predicted OCT4-pg4 protein contains NTD and POU domain, but lacks a large part of the C-terminal domain. Vertical lines within the depicted structures of OCT4 pseudogenes indicate the position of point mutations which have changed the amino acid sequences of the predicted proteins and B. Western blotting in different human cell types transfected with OCT4-Pg1, OCT4-Pg3 and OCT4-Pg4 expression vectors. The NCCIT and NT2 cell lines were used as positive controls for OCT4A, while U-87MG was used as a negative control for OCT4A and OCT4 pseudogenes. Using the sc-5279 antibody, we detected OCT4A protein exclusively in NCCIT and NTERA-2 cell lines but did not detect OCT4 pseudogenes in the cells which expressed them at the transcript level. Note that the internal control b-actin protein is detectable at similar intensities in all examined cell lines.

Article Snippet: In this experimental study, 17 human cell lines were mainly provided by Pasteur Institute of Iran and Research Institute of Avicenna, included two human embryonic carcinoma cell lines, one normal fibroblast cell line (HS-5) and HEK293 (embryonic kidney) and 13 human tumor cell lines, namely U87MG and A172 (glioblastoma), Daoy (medulloblastoma), 1321N1 (brain astrocytoma), Jurkat (T-Cell lymphoma), Y79 (retinoblastoma), PC3 (prostate adenocarcinoma), Raji (Burkit’s lymphoma), Ovcar3 (ovary adenocarcinoma), HepG2 (hepatocellular carcinoma), MCF7 (breast adenocarcinoma), 5637 (urinary bladder carcinoma), HeLa (cervix adenocarcinoma), and NT2 and NCCIT (pluripotent embryonic carcinoma).

Techniques: Western Blot, Transfection, Expressing, Negative Control, Control