Journal: OncoTargets and Therapy

Article Title:

Antitumor Effect of miR-1294/Pyruvate Kinase M2 Signaling Cascade in Osteosarcoma Cells

doi: 10.2147/ott.s232718

Figure Lengend Snippet: Figure 4 PKM2 was a target of miR-1294 in osteosarcoma cells. (A) PKM2 expression in human mesenchymal stem cells (hMSCs) and five osteosarcoma cell lines (Saos-2, MG63, U2OS, HOS, and 143B) was assessed by real-time PCR. (B) The miR-1294 binding site predictions for PKM2 3ʹUTR by microRNA.org. (C) A dual luciferase reporter assay was performed to confirm the binding in 293T cells. (D and E) PKM2 expression in U2OS and HOS cells transfected with miR-1294 mimic by real-time PCR and Western blot assays. (F and G) PKM2 expression in 143B cells transfected with miR-1294 inhibitor by real-time PCR and Western blot assays. Data are shown as mean ± SD. ∗P < 0.05, ∗∗∗P < 0.001.

Article Snippet: After dewaxing with xylene and rehydration with gradient ethanol, the sections were subjected to antigen retrieval and incubation with one of the primary antibodies against c-Myc (1:200; Proteintech), cyclin D1 (1:200; Boster), MMP2; (1:200; Proteintech), MMP-9 (1:200; Proteintech), and PKM2 (1:200; Proteintech) overnight at 4°C.

Techniques: Expressing, Real-time Polymerase Chain Reaction, Binding Assay, Luciferase, Reporter Assay, Transfection, Western Blot

Journal: OncoTargets and Therapy

Article Title:

Antitumor Effect of miR-1294/Pyruvate Kinase M2 Signaling Cascade in Osteosarcoma Cells

doi: 10.2147/ott.s232718

Figure Lengend Snippet: Figure 5 Overexpression of PKM2 reversed the actions of miR-1294 upregulation on osteosarcoma cells. (A) Cell proliferation was evaluated by CCK-8 assay. (B) Cell apoptosis was examined by flow cytometry using Annexin Vand PI staining. (C) Cell migration was assessed by a wound-healing assay. (D) Cell invasiveness was determined by transwell assay. (E) Expression levels of c-Myc, cyclin D1, cleaved-caspase 3, MMP-2, and MMP-9 were texted by Western blotting. Data are shown as mean ± SD. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001.

Article Snippet: After dewaxing with xylene and rehydration with gradient ethanol, the sections were subjected to antigen retrieval and incubation with one of the primary antibodies against c-Myc (1:200; Proteintech), cyclin D1 (1:200; Boster), MMP2; (1:200; Proteintech), MMP-9 (1:200; Proteintech), and PKM2 (1:200; Proteintech) overnight at 4°C.

Techniques: Over Expression, CCK-8 Assay, Cytometry, Staining, Migration, Wound Healing Assay, Transwell Assay, Expressing, Western Blot

Journal: OncoTargets and Therapy

Article Title:

Antitumor Effect of miR-1294/Pyruvate Kinase M2 Signaling Cascade in Osteosarcoma Cells

doi: 10.2147/ott.s232718

Figure Lengend Snippet: Figure 6 Overexpression of miR-1294 inhibited tumorigenicity of osteosarcoma cells. (A, B) Tumor volume in nude mice after subcutaneous injection of ME63 and U2O2 cells with or without overexpression of miR-1294. (C) The expression of miR-1294 in tumor tissues was determined by real-time PCR. (D, E) The expression of PKM2, c-Myc, cyclin D1, MMP-2, and MMP-9 in tumor tissues was assessed using immunohistochemistry and Western blot analysis. Data are shown as mean ± SD from 6 mice/ group. ∗P < 0.05, ∗∗∗P < 0.001.

Article Snippet: After dewaxing with xylene and rehydration with gradient ethanol, the sections were subjected to antigen retrieval and incubation with one of the primary antibodies against c-Myc (1:200; Proteintech), cyclin D1 (1:200; Boster), MMP2; (1:200; Proteintech), MMP-9 (1:200; Proteintech), and PKM2 (1:200; Proteintech) overnight at 4°C.

Techniques: Over Expression, Injection, Expressing, Real-time Polymerase Chain Reaction, Immunohistochemistry, Western Blot

Journal: iScience

Article Title: Lamin chromatin binding is modulated by interactions of different LAP2α domains with lamins and chromatin

doi: 10.1016/j.isci.2024.110869

Figure Lengend Snippet:

Article Snippet: Anti-BANF1 M2 (mouse, monoclonal) , Novus Biologicals , Cat# H00008815-M07; RRID: AB_537085.

Techniques: Recombinant, Plasmid Preparation, Protease Inhibitor, Magnetic Beads, Extraction, Mutagenesis, Generated, Knock-Out, Double Knockout, Control, Software, Microscopy, Sonication, Pore Size