control plasmid (pcdna™3.1)  (Thermo Fisher)

Journal: BMC Musculoskeletal Disorders

Article Title: FGFR1 inhibits skeletal muscle atrophy associated with hindlimb suspension

doi: 10.1186/1471-2474-8-32

Figure Lengend Snippet: Ectopic expression of FGFR1 inhibits atrophy . β-galactosidase expression in muscle fibers from gastrocnemius muscles of mice co-transfected with a β-galactosidase expression vector and either a control (control) or FGFR1 expression plasmid DNA (FGFR1). Rows represent fibers from muscles of mice suspended for 7 d (7 d HS), 14 d (14 d HS), or reloaded for 7 d after 7 d HS (reloaded). The bar in the bottom right figure represents 50 μm.

Article Snippet: Control pcDNA 3.1 (+) plasmid DNA was purchased from Invitrogen (catalog no. V790-20, Carlsbad, CA).

Techniques: Expressing, Transfection, Plasmid Preparation

Journal: BMC Musculoskeletal Disorders

Article Title: FGFR1 inhibits skeletal muscle atrophy associated with hindlimb suspension

doi: 10.1186/1471-2474-8-32

Figure Lengend Snippet: Overexpression of FGFR1 inhibits muscle atrophy . Cross-sectional area (CSA) of fibers from gastrocnemius muscles of mice transfected with either control (control) or FGFR1 expression plasmid DNA (FGFR1), subjected to hindlimb suspension for 7 d (7 d HS), 14 d (14 d HS), reloaded for 7 d after 7 d of hindlimb suspension (7 d reloaded), or non-suspended controls (NS). Means (±SEM) bearing different letters differ significantly (P < 0.05).

Article Snippet: Control pcDNA 3.1 (+) plasmid DNA was purchased from Invitrogen (catalog no. V790-20, Carlsbad, CA).

Techniques: Over Expression, Transfection, Expressing, Plasmid Preparation

Journal: BMC Musculoskeletal Disorders

Article Title: FGFR1 inhibits skeletal muscle atrophy associated with hindlimb suspension

doi: 10.1186/1471-2474-8-32

Figure Lengend Snippet: Overexpression of skeletal muscle with FGFR1 increases protein synthesis . Incorporation of L-[4- 3 H] phenylalanine in gastrocnemius and soleus muscles of weight bearing or hindlimb suspended mice transfected with control (control) or FGFR1 expression plasmid DNA (FGFR1). Values represent back transformation of means (+SE/-SE) generated using a square root transformation. Values bearing different letters differ significantly ( P < 0.05).

Article Snippet: Control pcDNA 3.1 (+) plasmid DNA was purchased from Invitrogen (catalog no. V790-20, Carlsbad, CA).

Techniques: Over Expression, Transfection, Expressing, Plasmid Preparation, Transformation Assay, Generated

Journal: BMC Musculoskeletal Disorders

Article Title: FGFR1 inhibits skeletal muscle atrophy associated with hindlimb suspension

doi: 10.1186/1471-2474-8-32

Figure Lengend Snippet: Proteasome activity is unaffected by overexpression of FGFR1 in muscle subjected to hindlimb suspension (HS) . Luminescence of extracts from the gastrocnemius and soleus muscles of mice transfected with either control (control) or FGFR1 expression plasmid DNA (FGFR1), subjected to either hindlimb suspension (HS), or non-suspended controls (NS). Data are expressed as luciferase dependent light units of Ub-Fl normalized to renilla luciferase dependent light units. Means (±SEM) bearing different letters differ significantly (P < 0.05).

Article Snippet: Control pcDNA 3.1 (+) plasmid DNA was purchased from Invitrogen (catalog no. V790-20, Carlsbad, CA).

Techniques: Activity Assay, Over Expression, Transfection, Expressing, Plasmid Preparation, Luciferase

Journal: BMC Musculoskeletal Disorders

Article Title: FGFR1 inhibits skeletal muscle atrophy associated with hindlimb suspension

doi: 10.1186/1471-2474-8-32

Figure Lengend Snippet: Downstream detection of activated FGF signaling . Activity of mouse osteocalcin gene 2 promoter (mOG2)-luciferase reporter gene in gastrocnemius and soleus muscles of mice transfected with either control (control) or FGFR1 expression plasmid DNA (FGFR1). Data are expressed as luciferase dependent light units normalized to renilla luciferase dependent light units (mOG2/Fl). Means (±SEM) bearing different letters differ significantly ( P < 0.01).

Article Snippet: Control pcDNA 3.1 (+) plasmid DNA was purchased from Invitrogen (catalog no. V790-20, Carlsbad, CA).

Techniques: Activity Assay, Luciferase, Transfection, Expressing, Plasmid Preparation

Journal: Bone & Joint Research

Article Title: CircSCAPER contributes to IL-1β-induced osteoarthritis in vitro via miR-140-3p/EZH2 axis

doi: 10.1302/2046-3758.112.BJR-2020-0482.R2

Figure Lengend Snippet: MicroRNA (miR)-140-3p protects chondrocytes against interleukin (IL)-1β-induced dysfunction through enhancer of zeste homolog 2 (EZH2). a) to e) C28/I2 cells were transfected with miR-NC, miR-140-3p, miR-140-3p+pcDNA, or miR-140-3p+EZH2, followed by treatment with IL-1β (10 ng/l) for 24 hours. a) Western blot analysis of EZH2 protein level in cells. b) Western blot analysis of matrix metallopeptidase 13 (MMP-13), a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-5, and collagen II protein levels in cells. c) Cell counting kit-8 (CCK-8) assay for cell proliferation. d) Flow cytometry for cell apoptosis. e) Detection of B-cell lymphoma-2 (Bcl-2), Bcl-2-associated X (Bax), and cleaved caspase 3 protein levels in cells using Western blot. One-way analysis of variance (ANOVA), *p < 0.05. All of the experiments were performed in triplicate.

Article Snippet: The miR-140-3p mimic or inhibitor (miR-140-3p or in-miR-140-3p) and their negative control (miR-NC or in-miR-NC), circSCAPER-specific small interfering RNA (siRNA) (si-circSCAPER) and negative control (si-NC), pCD25-ciR-circSCAPER overexpression plasmid (circSCAPER) and plasmid empty control (NC), pcDNA 3.1-EZH2 overexpression plasmid (EZH2) and plasmid empty control (pcDNA) were obtained from Genepharma (China).

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

Journal: Bone & Joint Research

Article Title: CircSCAPER contributes to IL-1β-induced osteoarthritis in vitro via miR-140-3p/EZH2 axis

doi: 10.1302/2046-3758.112.BJR-2020-0482.R2

Figure Lengend Snippet: CircSCAPER can activate the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT) pathway through the microRNA (miR)-140-3p/enhancer of zeste homolog 2 (EZH2) axis. a) Western blot analysis of p-AKT and p-PI3K levels in C28/I2 cells transfected with circSCAPER-specific small interfering RNA (siRNA) (si-circSCAPER), si-negative control (NC), si-circSCAPER+in-miR-NC, or si-circSCAPER+in-miR-140-3p in the presence of interleukin (IL)-1β. b) Western blot analysis of p-AKT and p-PI3K levels in C28/I2 cells transfected with miR-NC, miR-140-3p, miR-140-3p+pcDNA, or miR-140-3p+EZH2 in the presence of IL-1β. One-way analysis of variance, *p < 0.05. All of the experiments were performed in triplicate.

Article Snippet: The miR-140-3p mimic or inhibitor (miR-140-3p or in-miR-140-3p) and their negative control (miR-NC or in-miR-NC), circSCAPER-specific small interfering RNA (siRNA) (si-circSCAPER) and negative control (si-NC), pCD25-ciR-circSCAPER overexpression plasmid (circSCAPER) and plasmid empty control (NC), pcDNA 3.1-EZH2 overexpression plasmid (EZH2) and plasmid empty control (pcDNA) were obtained from Genepharma (China).

Techniques: Western Blot, Transfection, Small Interfering RNA, Negative Control

Journal: Frontiers in Genetics

Article Title: Long Non-coding RNA FGD5-AS1 Regulates Cancer Cell Proliferation and Chemoresistance in Gastric Cancer Through miR-153-3p/CITED2 Axis

doi: 10.3389/fgene.2020.00715

Figure Lengend Snippet: Overexpressing CITED2 also restored cancer cell proliferation and chemoresistance in gastric cancer cells with FGD5-AS1 downregulation. (A) In sh-FGD5AS1-transduced SGC-7901 and MKN-28 cells, a mammalian expression plasmid containing the whole cDNA sequence of human CITED2 (pc/CITED2), or an empty control pcDNA 3.1/+ plasmid (pc/C), was transfected into cells. After 48 h, qRT-PCR was carried out to verify the transfection efficiency (* P < 0.05). (B) A 5-day MTT assay was carried out to compare the proliferation rates between SGC-7901 and MKN-28 cells infected with sh-FFG5AS1 and pc/C, and those infected with sh-FFG5AS1 and pc/CITED2 (** P < 0.05). (C) In double-infected SGC-7901 and MKN-28 cells, 5-FU chemoresistance was compared between cells infected with sh-FFG5AS1 and pc/C, and those infected with sh-FFG5AS1 and pc/CITED2 (** P < 0.05).

Article Snippet: A mammalian expression plasmid (pcDNA 3.1/+) containing the whole cDNA sequence of human CITED2 (pc/CITED2) and an empty control pcDNA 3.1/+ plasmid (pc/C) were purchased from RiboBio (RiboBio, Guangzhou, China).

Techniques: Expressing, Plasmid Preparation, Sequencing, Transfection, Quantitative RT-PCR, MTT Assay, Infection