Journal: Frontiers in Oncology

Article Title: A Schistosoma japonicum MicroRNA Exerts Antitumor Effects Through Inhibition of Both Cell Migration and Angiogenesis by Targeting PGAM1

doi: 10.3389/fonc.2021.652395

Figure Lengend Snippet: PGAM1 as target of sja-miR-61. (A) A schematic diagram representing the wild-type or mutant 3’ untranslated region (UTR) targeting sites of murine Pgam1 and human PGAM1 genes. (B) A dual-luciferase reporter assay was used to measure the activity of the reporter gene, and the firefly luciferase activity was normalized to renilla luciferase activity. (C, D) The protein levels of murine PGAM1 (C) and human PGAM1 (D) were measured using Western blotting in the hepatoma cells transfected with sja-miR-61 mimics or NC mimics, respectively. Data are presented as the mean ± SD, n = 3, ** p < 0.01.

Article Snippet: Then the membrane was blocked with 5% bovine serum albumin (BSA) in Tris-buffered saline with Tween (TBST) for 2 h at room temperature, followed by incubated overnight with primary antibodies against PGAM1 (1:500 dilution, Proteintech, China), GAPDH (1:1000 dilution, Beyotime, China).

Techniques: Mutagenesis, Luciferase, Reporter Assay, Activity Assay, Western Blot, Transfection

Journal: Frontiers in Oncology

Article Title: A Schistosoma japonicum MicroRNA Exerts Antitumor Effects Through Inhibition of Both Cell Migration and Angiogenesis by Targeting PGAM1

doi: 10.3389/fonc.2021.652395

Figure Lengend Snippet: Knockdown of PGAM1 inhibits cell migration of hepa1-6 and HepG2 cells in vitro. (A, B) Hepa1-6 and HepG2 cells were transfected with PGAM1 siRNA and negative control (NC) siRNA, respectively, and 48 h later, the expression of PGAM1 was determined using qRT-PCR and Western blotting (A) . Cell migration was evaluated using transwell inserts without matrigel coating (B) . Data are presented as the mean ± SD, n = 3, ** p < 0.01.

Article Snippet: Then the membrane was blocked with 5% bovine serum albumin (BSA) in Tris-buffered saline with Tween (TBST) for 2 h at room temperature, followed by incubated overnight with primary antibodies against PGAM1 (1:500 dilution, Proteintech, China), GAPDH (1:1000 dilution, Beyotime, China).

Techniques: Knockdown, Migration, In Vitro, Transfection, Negative Control, Expressing, Quantitative RT-PCR, Western Blot

Journal: Frontiers in Oncology

Article Title: A Schistosoma japonicum MicroRNA Exerts Antitumor Effects Through Inhibition of Both Cell Migration and Angiogenesis by Targeting PGAM1

doi: 10.3389/fonc.2021.652395

Figure Lengend Snippet: Effect of restoration of PGAM1 expression on the sja-miR-61-mediated effects in vitro . (A–D) Cells were transfected with NC mimics, sja-miR-61 mimics, NC mimics + pcDNA3.1(+) vector, sja-miR-61 mimics + pcDNA3.1(+)-PGAM1, respectively, and 48 h later, the expression of PGAM1 was determined using Western blotting (A, B) . Cell migration was evaluated using transwell inserts without matrigel coating (C, D) . Mean ± SD, n = 3, **p < 0.01.

Article Snippet: Then the membrane was blocked with 5% bovine serum albumin (BSA) in Tris-buffered saline with Tween (TBST) for 2 h at room temperature, followed by incubated overnight with primary antibodies against PGAM1 (1:500 dilution, Proteintech, China), GAPDH (1:1000 dilution, Beyotime, China).

Techniques: Expressing, In Vitro, Transfection, Plasmid Preparation, Western Blot, Migration

Journal: Frontiers in Oncology

Article Title: A Schistosoma japonicum MicroRNA Exerts Antitumor Effects Through Inhibition of Both Cell Migration and Angiogenesis by Targeting PGAM1

doi: 10.3389/fonc.2021.652395

Figure Lengend Snippet: Sja-miR-61-mediated inhibition of migration and tube formation of HUVEC cells by down-regulating PGAM1 in vitro . (A, B) HUVEC cells were transfected with sja-miR-61 mimics and NC mimics (A) , PGAM1 siRNA and NC siRNA (B) , respectively, and 48 h later, the expression of PGAM1 was determined using qRT-PCR. Cell migration was evaluated using transwell inserts without matrigel coating (C) . (D, E) The sja-miR-61 mimics within the transfected cells were measured by qRT-PCR at day 0 (i.e. before inoculation) and 8 post inoculation, using U6 as the internal control, (D) for Hepa1-6 cells, (E) for HepG2 cells. (F) The sja-miR-61 mimics were detected by qRT-PCR in HUVEC cells cultured with conditioned medium (CM) derived from the cultivation of HepG2 cells transfected with sja-miR-61 mimics or NC mimics. Mean ± SD, n = 3, **p < 0.01.

Article Snippet: Then the membrane was blocked with 5% bovine serum albumin (BSA) in Tris-buffered saline with Tween (TBST) for 2 h at room temperature, followed by incubated overnight with primary antibodies against PGAM1 (1:500 dilution, Proteintech, China), GAPDH (1:1000 dilution, Beyotime, China).

Techniques: Inhibition, Migration, In Vitro, Transfection, Expressing, Quantitative RT-PCR, Control, Cell Culture, Derivative Assay

Journal: Cell & Bioscience

Article Title: SARS-CoV-2 NSP13 helicase suppresses interferon signaling by perturbing JAK1 phosphorylation of STAT1

doi: 10.1186/s13578-022-00770-1

Figure Lengend Snippet: SARS-CoV-2 NSP13 suppresses type I and type II IFN signaling. A NSP13 suppresses IFN-β- and IFN-γ- induced ISRE and GAS promoter activity. HEK293T cells were transfected with pISRE-Luc or pGAS-Luc, SV40 Renilla luciferase and increasing doses of NSP13 plasmid (200, 400 and 600 ng). At 24 h post transfection, cells were stimulated by 1000 U/mL of IFN-β or 100 ng/mL of IFN-γ. Dual luciferase activity was measured 24 h post IFN treatment. B – G Suppression of IFN-β and IFN-γ signaling by NSP13. A549 cells were either mock transfected or transfected with NSP13 plasmid. At 26 h post transfection, cells were stimulated by 1000 U/mL IFN-β or 100 ng/mL IFN-γ for 6 h. ISG transcripts were analyzed by RT-qPCR. Results were representative of three independent experiments. The statistical significance of the differences between the indicated groups was evaluated by one-tailed Student t test for unpaired samples with equal variance. *P < 0.05. **P < 0.01. ***P < 0.001

Article Snippet: Luciferase reporter constructs pISRE-Luc and pGAS-Luc were from Promega.

Techniques: Activity Assay, Transfection, Luciferase, Plasmid Preparation, Quantitative RT-PCR, One-tailed Test