agps shrna plasmid  (Santa Cruz Biotechnology)

Journal: Oncology Letters

Article Title: Tudor-staphylococcal nuclease regulates the expression and biological function of alkylglycerone phosphate synthase via nuclear factor-κB and microRNA-127 in human glioma U87MG cells

doi: 10.3892/ol.2018.8484

Figure Lengend Snippet: Tudor-SN silencing reduces the proliferation and migration in human glioma U87MG cells. (A) The cell proliferation assay demonstrated that the potential for proliferation was reduced by Tudor-SN silencing in human glioma U87MG cells. (B) The Transwell assay indicated that the potential for migration was reduced by Tudor-SN silencing in human glioma U87MG cells. *P≤0.05, compared with control group. AGPS, alkylglycerone phosphate synthase; Tudor-SN, Tudor-staphylococcal nuclease; shRNA, short hairpin RNA.

Article Snippet: A total of 3×10 5 cells/well were seeded onto a 6-well plate and cultured at 37°C for 24 h. A total of 2.5 μg AGPS shRNA plasmid, 2.5 μg Tudor-SN shRNA plasmid (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), 2.5 μg NF-κB p65 expression plasmid (Santa Cruz Biotechnology, Inc.) and 2.5 μg miR-127 siRNA plasmid (OBIO Biotechnology, Inc., Shanghai, China) were transfected using GeneJuice ® (Merck KGaA, Darmstadt, Germany), according to the manufacturer's protocol, for another 6 h. Fresh Dulbecco's modified Eagle's medium (Corning Life Sciences, Manassas, VA, USA) was then added and the cells were harvested after 72 h for all experiments.

Techniques: Migration, Proliferation Assay, Transwell Assay, Control, shRNA

Journal: Oncology Letters

Article Title: Tudor-staphylococcal nuclease regulates the expression and biological function of alkylglycerone phosphate synthase via nuclear factor-κB and microRNA-127 in human glioma U87MG cells

doi: 10.3892/ol.2018.8484

Figure Lengend Snippet: AGPS silencing regulates the expression of circRNAs and lncRNAs in human glioma U87MG cells. The reverse transcription-quantitative polymerase chain reaction assay demonstrated that AGPS silencing regulated the expression of circRNAs (circUBAP2, circZNF292 and circHIPK3) and lncRNAs (H19, CCAT1 and HULC) in human glioma U87MG cells. *P≤0.05, compared with control group. AGPS, alkylglycerone phosphate synthase; shRNA, short hairpin RNA; circ, circular; UBAP, ubiquitin-associated protein 2; ZNF292, zinc finger protein 292; HIPK3, homeodomain-interacting protein kinase 3; H19, H19 imprinted maternally expressed transcript (non-protein coding); CCAT1, colon cancer-associated transcript 1 (non-protein coding); HULC, hepatocellular carcinoma upregulated long non-coding RNA.

Article Snippet: A total of 3×10 5 cells/well were seeded onto a 6-well plate and cultured at 37°C for 24 h. A total of 2.5 μg AGPS shRNA plasmid, 2.5 μg Tudor-SN shRNA plasmid (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), 2.5 μg NF-κB p65 expression plasmid (Santa Cruz Biotechnology, Inc.) and 2.5 μg miR-127 siRNA plasmid (OBIO Biotechnology, Inc., Shanghai, China) were transfected using GeneJuice ® (Merck KGaA, Darmstadt, Germany), according to the manufacturer's protocol, for another 6 h. Fresh Dulbecco's modified Eagle's medium (Corning Life Sciences, Manassas, VA, USA) was then added and the cells were harvested after 72 h for all experiments.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, shRNA, Ubiquitin Proteomics

Journal: Oncology Letters

Article Title: Tudor-staphylococcal nuclease regulates the expression and biological function of alkylglycerone phosphate synthase via nuclear factor-κB and microRNA-127 in human glioma U87MG cells

doi: 10.3892/ol.2018.8484

Figure Lengend Snippet: Tudor-SN silencing decreases the expression of AGPS via NF-κB and miR-127 in human glioma U87MG cells. (A) The western blotting assay indicated that Tudor-SN silencing decreased the protein expression of AGPS, and that the expression of AGPS was partially recovered via NF-κB and miR-127 retrieval experiments. (B) The reverse transcription-polymerase chain reaction assay demonstrated that Tudor-SN silencing decreased the mRNA expression of AGPS, and that the mRNA expression of AGPS was partially recovered via NF-κB and miR-127 retrieval experiments. *P≤0.05, compared with control group; #P≤0.05, compared with Tudor-SN silencing group. AGPS, alkylglycerone phosphate synthase; Tudor-SN, Tudor-staphylococcal nuclease; NF, nuclear factor; mir, microRNA; shRNA, short hairpin RNA.

Article Snippet: A total of 3×10 5 cells/well were seeded onto a 6-well plate and cultured at 37°C for 24 h. A total of 2.5 μg AGPS shRNA plasmid, 2.5 μg Tudor-SN shRNA plasmid (Santa Cruz Biotechnology, Inc., Dallas, TX, USA), 2.5 μg NF-κB p65 expression plasmid (Santa Cruz Biotechnology, Inc.) and 2.5 μg miR-127 siRNA plasmid (OBIO Biotechnology, Inc., Shanghai, China) were transfected using GeneJuice ® (Merck KGaA, Darmstadt, Germany), according to the manufacturer's protocol, for another 6 h. Fresh Dulbecco's modified Eagle's medium (Corning Life Sciences, Manassas, VA, USA) was then added and the cells were harvested after 72 h for all experiments.

Techniques: Expressing, Western Blot, Reverse Transcription, Polymerase Chain Reaction, Control, shRNA

Journal: Oncology Letters

Article Title: Effect of alkylglycerone phosphate synthase on the expression profile of circRNAs in the human thyroid cancer cell line FRO

doi: 10.3892/ol.2018.8356

Figure Lengend Snippet: Primer sequences.

Article Snippet: FRO cells were cultured in a 6-well plate for 24 h, and then transfected with an AGPS short hairpin (sh)RNA plasmid (sc-94310-SH; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) (AGPS sh group) or with an AGPS clustered regularly interspaced short palindromic repeats-associated protein-9 nuclease knockout (KO) plasmid (sc-404604; Santa Cruz Biotechnology, Inc.) (AGPS KO group), according to the manufacturer's protocol.

Techniques: Sequencing

Journal: Oncology Letters

Article Title: Effect of alkylglycerone phosphate synthase on the expression profile of circRNAs in the human thyroid cancer cell line FRO

doi: 10.3892/ol.2018.8356

Figure Lengend Snippet: Expression of AGPS in thyroid cancer cell lines. AGPS expression in the thyroid cancer cell lines TPC-1, FTC-133 and FRO was evaluated using reverse transcription-quantitative polymerase chain reaction analysis. *P<0.05 vs. TPC-1. AGPS, alkylglycerone phosphate synthase.

Article Snippet: FRO cells were cultured in a 6-well plate for 24 h, and then transfected with an AGPS short hairpin (sh)RNA plasmid (sc-94310-SH; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) (AGPS sh group) or with an AGPS clustered regularly interspaced short palindromic repeats-associated protein-9 nuclease knockout (KO) plasmid (sc-404604; Santa Cruz Biotechnology, Inc.) (AGPS KO group), according to the manufacturer's protocol.

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Effect of alkylglycerone phosphate synthase on the expression profile of circRNAs in the human thyroid cancer cell line FRO

doi: 10.3892/ol.2018.8356

Figure Lengend Snippet: Heatmap of circRNA expression data. The different expressions of circRNAs were depicted in the heatmap. Green represents a decreased expression of circRNAs and red represents an increased expression of circRNAs. Cluster analysis demonstrated that AGPS sh and AGPS KO could induce the alteration in circRNAs expression in FRO cells. circRNA, circular RNA; AGPS, alkylglycerone phosphate synthase; sh, short hairpin; KO, knockout.

Article Snippet: FRO cells were cultured in a 6-well plate for 24 h, and then transfected with an AGPS short hairpin (sh)RNA plasmid (sc-94310-SH; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) (AGPS sh group) or with an AGPS clustered regularly interspaced short palindromic repeats-associated protein-9 nuclease knockout (KO) plasmid (sc-404604; Santa Cruz Biotechnology, Inc.) (AGPS KO group), according to the manufacturer's protocol.

Techniques: Expressing, Knock-Out

Journal: Oncology Letters

Article Title: Effect of alkylglycerone phosphate synthase on the expression profile of circRNAs in the human thyroid cancer cell line FRO

doi: 10.3892/ol.2018.8356

Figure Lengend Snippet: Comparison of differentially expressed circRNAs. (A) Differentially expressed circRNAs among the different groups were identified by scatter plots. The orange points represent >2-fold upregulation, whereas the blue points represent >2-fold downregulation. (B) Differences in circRNA expression among the three groups. circRNA, circular RNA; AGPS, alkylglycerone phosphate synthase; sh, short hairpin; KO, knockout.

Article Snippet: FRO cells were cultured in a 6-well plate for 24 h, and then transfected with an AGPS short hairpin (sh)RNA plasmid (sc-94310-SH; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) (AGPS sh group) or with an AGPS clustered regularly interspaced short palindromic repeats-associated protein-9 nuclease knockout (KO) plasmid (sc-404604; Santa Cruz Biotechnology, Inc.) (AGPS KO group), according to the manufacturer's protocol.

Techniques: Comparison, Expressing, Knock-Out

Journal: Oncology Letters

Article Title: Effect of alkylglycerone phosphate synthase on the expression profile of circRNAs in the human thyroid cancer cell line FRO

doi: 10.3892/ol.2018.8356

Figure Lengend Snippet: Venn diagrams of differentially expressed circular RNAs in the three groups. The Venn diagrams represent the shared modified genes between groups: AGPS_sh vs. control; AGPS_KO vs. control; and AGPS_sh vs. AGPS_KO, in FRO cell lines. AGPS, alkylglycerone phosphate synthase; sh, short hairpin; KO, knockout.

Article Snippet: FRO cells were cultured in a 6-well plate for 24 h, and then transfected with an AGPS short hairpin (sh)RNA plasmid (sc-94310-SH; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) (AGPS sh group) or with an AGPS clustered regularly interspaced short palindromic repeats-associated protein-9 nuclease knockout (KO) plasmid (sc-404604; Santa Cruz Biotechnology, Inc.) (AGPS KO group), according to the manufacturer's protocol.

Techniques: Modification, Control, Knock-Out

Journal: Oncology Letters

Article Title: Effect of alkylglycerone phosphate synthase on the expression profile of circRNAs in the human thyroid cancer cell line FRO

doi: 10.3892/ol.2018.8356

Figure Lengend Snippet: Top 50 differentially expressed circRNAs in AGPS sh and AGPS KO human thyroid cancer FRO cells.

Article Snippet: FRO cells were cultured in a 6-well plate for 24 h, and then transfected with an AGPS short hairpin (sh)RNA plasmid (sc-94310-SH; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) (AGPS sh group) or with an AGPS clustered regularly interspaced short palindromic repeats-associated protein-9 nuclease knockout (KO) plasmid (sc-404604; Santa Cruz Biotechnology, Inc.) (AGPS KO group), according to the manufacturer's protocol.

Techniques: Control

Journal: Oncology Letters

Article Title: Effect of alkylglycerone phosphate synthase on the expression profile of circRNAs in the human thyroid cancer cell line FRO

doi: 10.3892/ol.2018.8356

Figure Lengend Snippet: Validation of differentially expressed circRNAs using RT-qPCR. The top 10 circRNAs with increased and decreased expression in the AGPS KO group compared with the control group by RT-qPCR were identified. *P<0.05 vs. control. circRNA, circular RNA; RT-qPCR, reverse transcription-quantitative polymerase chain reaction; AGPS, alkylglycerone phosphate synthase; sh, short hairpin; KO, knockout; hsa, Homo sapiens .

Article Snippet: FRO cells were cultured in a 6-well plate for 24 h, and then transfected with an AGPS short hairpin (sh)RNA plasmid (sc-94310-SH; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) (AGPS sh group) or with an AGPS clustered regularly interspaced short palindromic repeats-associated protein-9 nuclease knockout (KO) plasmid (sc-404604; Santa Cruz Biotechnology, Inc.) (AGPS KO group), according to the manufacturer's protocol.

Techniques: Biomarker Discovery, Quantitative RT-PCR, Expressing, Control, Reverse Transcription, Real-time Polymerase Chain Reaction, Knock-Out

Journal: Oncology Letters

Article Title: Effect of alkylglycerone phosphate synthase on the expression profile of circRNAs in the human thyroid cancer cell line FRO

doi: 10.3892/ol.2018.8356

Figure Lengend Snippet: GO analysis of AGPS silencing in thyroid cancer FRO cells. GO analysis was performed on the up- and downregulated genes identified in the AGPS sh, AGPS KO and control thyroid cancer FRO cells. The y-axis represented gene ontology entries, the x-axis represented -log10 (P-value). -log10 (P-value) >2 was considered to indicate a significant threshold. (A) Upregulation and (B) downregulation of MF, BP and CC by GO analysis of AGPS silencing in FRO cells compared with control thyroid cancer FRO cells. (C) Upregulation and (D) downregulation of MF, BP and CC by GO analysis of AGPS silencing in FRO cells compared with AGPS KO in thyroid cancer FRO cells. (E) Upregulation and (F) downregulation of MF, BP and CC by GO analysis of AGPS KO compared with control thyroid cancer FRO cells. GO, Gene Ontology; AGPS, alkylglycerone phosphate synthase; sh, short hairpin; KO, knockout; MF, molecular function; BP, biological process; CC, cellular component.

Article Snippet: FRO cells were cultured in a 6-well plate for 24 h, and then transfected with an AGPS short hairpin (sh)RNA plasmid (sc-94310-SH; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) (AGPS sh group) or with an AGPS clustered regularly interspaced short palindromic repeats-associated protein-9 nuclease knockout (KO) plasmid (sc-404604; Santa Cruz Biotechnology, Inc.) (AGPS KO group), according to the manufacturer's protocol.

Techniques: Control, Knock-Out

Journal: Oncology Letters

Article Title: Effect of alkylglycerone phosphate synthase on the expression profile of circRNAs in the human thyroid cancer cell line FRO

doi: 10.3892/ol.2018.8356

Figure Lengend Snippet: KEGG pathway enrichment analysis of AGPS silencing in thyroid cancer FRO cells. KEGG pathway enrichment analysis is represented by histograms and scatter diagrams. Orange represents the pathway of upregulation gene enrichment, whereas blue represents the downregulated pathway of gene enrichment. The left and right sides of the scatter diagrams correspond to the downregulated and upregulated gene enrichment pathways, respectively. The x-axes represent the proportion of enriched genes, whereas the y-axes contain the name of the KEGG pathway. If up- or downregulated genes were enriched in a certain pathway, this pathway was represented as a point in the graph, the color of which indicated a significant P-value, whereas its size represented the number of differentially expressed genes in that particular pathway. Effect of regulatory function and signaling pathway by KEGG pathway enrichment analysis of AGPS silencing compared with (A) control and (B) KO thyroid cancer FRO cells. (C) Effect of regulatory function and signaling pathway by KEGG pathway enrichment analysis of AGPS KO compared with control thyroid cancer FRO cells. KEGG, Kyoto Encyclopedia of Genes and Genomes; AGPS, alkylglycerone phosphate synthase; KO, knockout; hsa, Homo sapiens .

Article Snippet: FRO cells were cultured in a 6-well plate for 24 h, and then transfected with an AGPS short hairpin (sh)RNA plasmid (sc-94310-SH; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) (AGPS sh group) or with an AGPS clustered regularly interspaced short palindromic repeats-associated protein-9 nuclease knockout (KO) plasmid (sc-404604; Santa Cruz Biotechnology, Inc.) (AGPS KO group), according to the manufacturer's protocol.

Techniques: Control, Knock-Out