Journal: Cell cycle (Georgetown, Tex.)

Article Title: TGF-β is associated with poor prognosis and promotes osteosarcoma progression via PI3K/Akt pathway activation.

doi: 10.1080/15384101.2020.1805552

Figure Lengend Snippet: Figure 1. TGF-β expression in osteosarcoma specimens and cell lines. (a) Immunohistochemical staining was used to analyze TGF-β expression in both osteosarcoma specimens and normal adjacent tissues. (b) Survival analysis was performed using the Kaplan-Meier method. Data are presented as mean ± SEM of three independent experiments. (c) TGF-β protein levels in osteosarcoma cell lines (*P < 0.05). (d) TGF-β mRNA expression in osteosarcoma cell lines (*P < 0.05).

Article Snippet: Osteosarcoma spheroids were cultured in a specialized growth medium (Celprogen Inc, Torrance, CA, USA) containing 1% N2 supplement (Invitrogen), 2% B27 supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen), and 1% antimycotic (Invitrogen).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Cell cycle (Georgetown, Tex.)

Article Title: TGF-β is associated with poor prognosis and promotes osteosarcoma progression via PI3K/Akt pathway activation.

doi: 10.1080/15384101.2020.1805552

Figure Lengend Snippet: Figure 2. Small interfering (si) RNA targeting of TGF-β (si-TGF-β) suppresses proliferation and promotes apoptosis in osteosarcoma. (a) RT-qPCR was employed to assess the knockdown efficiency of si-TGF-β, *P < 0.05 vs. the si-control group. (b) Western blotting was used to measure the knockdown efficiency of si-TGF-β, *P < 0.05 vs. the si-control group. (c)The MTT assay was used to assess the viability of U2OS and MG-63 cells transfected with si-control or si-TGF-β, *P < 0.05 vs. the si-control group. (d and e) The colony- forming ability of U2OS and MG-63 cells transfected with si-control or si-TGF -β, *P < 0.05 vs. the si-control group. (f and g) Flow cytometry analysis of the cell cycle phase distribution of U2OS and MG-63 cells transfected with si-control or si-TGF – β, *P < 0.05 vs. the si-control group. (h and i) The rate of apoptosis in U2OS and MG-63 cells transfected with si-TGF-β, *P < 0.05 vs. the si-control group. (j and k) TGF-β knockdown increases the expression of cleaved PARP, caspase-3, and BAX, *P < 0.05 vs. the si-control group.

Article Snippet: Osteosarcoma spheroids were cultured in a specialized growth medium (Celprogen Inc, Torrance, CA, USA) containing 1% N2 supplement (Invitrogen), 2% B27 supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen), and 1% antimycotic (Invitrogen).

Techniques: Quantitative RT-PCR, Knockdown, Control, Western Blot, MTT Assay, Transfection, Flow Cytometry, Expressing

Journal: Cell cycle (Georgetown, Tex.)

Article Title: TGF-β is associated with poor prognosis and promotes osteosarcoma progression via PI3K/Akt pathway activation.

doi: 10.1080/15384101.2020.1805552

Figure Lengend Snippet: Figure 4. TGF-β enhances osteosarcoma cell stemness and increases the proportion of CD133+ cells. (a and b) Spheroid formation assay for U2OS and MG-63 cells transfected with si-TGF-β. Representative images (left panel) and statistical measurements (right panel). Representative micrographs of formed spheres were analyzed in cells treated with si-TGF-β or si-control. Scale bar, 100 μm. *P < 0.05 vs. the si-control group. (c and d) The percentage of CD133+ cells in U2OS- and MG-63-derived spheroids was analyzed by flow cytometry.*P < 0.05 vs. the si-control group. (e and f) Stem cell-associated protein expression was measured by western blot. *P < 0.05 vs. the si-control group.

Article Snippet: Osteosarcoma spheroids were cultured in a specialized growth medium (Celprogen Inc, Torrance, CA, USA) containing 1% N2 supplement (Invitrogen), 2% B27 supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen), and 1% antimycotic (Invitrogen).

Techniques: Tube Formation Assay, Transfection, Control, Derivative Assay, Flow Cytometry, Expressing, Western Blot

Journal: Cell cycle (Georgetown, Tex.)

Article Title: TGF-β is associated with poor prognosis and promotes osteosarcoma progression via PI3K/Akt pathway activation.

doi: 10.1080/15384101.2020.1805552

Figure Lengend Snippet: Figure 5. TGF-β regulates the PI3K/mTOR signaling pathway in osteosarcoma cells. (a) p-PI3K and p-Akt expression was measured by western blot, P < 0.05 vs. the si-control group. (b and c) The colony-forming ability of si-TGF-β-transfected U2OS and MG-63 cells with or without LY294002 treatment. P < 0.05 vs. the si-control group. (d and e) Transwell assays were used to assess the invasive ability of U2OS and MG-63 cells, P < 0.05 vs. the si-control group. (f and g) A wound-healing assay was performed to measure the migratory ability of U2OS and MG-63 cells, P < 0.05 vs. the si-control group. (h and i) Spheroid formation assay for U2OS/MG-63 cells transfected with si-TGF-β. Representative images (left panel) and statistical measurement (right panel). Representative micrographs of formed spheres were analyzed in cells treated with si-TGF-β or si-control with or without LY294002 treatment. Scale bar, 100 μm. *P < 0.05 vs. the si-control group. (j and k) The percentage of CD133+ cells in U2OS cell- and MG-63 cell-derived spheroids was analyzed by flow cytometry. *P < 0.05 vs. the si-control group.

Article Snippet: Osteosarcoma spheroids were cultured in a specialized growth medium (Celprogen Inc, Torrance, CA, USA) containing 1% N2 supplement (Invitrogen), 2% B27 supplement (Invitrogen), 20 ng/ml human platelet growth factor (Sigma-Aldrich), 100 ng/ml epidermal growth factor (Invitrogen), and 1% antimycotic (Invitrogen).

Techniques: Expressing, Western Blot, Control, Transfection, Wound Healing Assay, Tube Formation Assay, Derivative Assay, Flow Cytometry

Journal: Cells

Article Title: Ischemic Stroke Risk Associated with Mitochondrial Haplogroup F in the Asian Population

doi: 10.3390/cells9081885

Figure Lengend Snippet: HIF-1α expressions in different cybrids under hypoxia-ischemia. Comparison of HIF-1α expression under hypoxic-ischemic conditions in cybrids harboring common mitochondrial haplogroups found in the ethnic Chinese population (B4, B5, D4, D5, F1, F2, N9). The 143B cybrid was used to represent the Caucasian population. Actin was used as a loading control. Data represent the mean ± SD of at least three independent experiments ( # p < 0.005). N was used to represent normoxic and H to represent the hypoxic condition for 24 h.

Article Snippet: MtDNA-depleted ρ0 cells were established by the treatment of human osteosarcoma 143B cells (BCRC 60439) (Bioresource Collection and Research Center, Taipei, Taiwan) with ethidium bromide for three months, and illustration of their mtDNA-less cellular status by RT-PCR has been previously described [ , ].

Techniques: Comparison, Expressing, Control

Journal: PLoS ONE

Article Title: The Coordinated P53 and Estrogen Receptor Cis-Regulation at an FLT1 Promoter SNP Is Specific to Genotoxic Stress and Estrogenic Compound

doi: 10.1371/journal.pone.0010236

Figure Lengend Snippet: To avoid any contribution of the endogenous p53 protein, gene reporter assays were performed in a p53-null, osteosarcoma-derived SaOS2 cell line. (A) Cells were transiently co-transfected with the FLT1-C or FLT1-T reporter constructs along with an overexpression vector for p53, p73β or p63γ as well as the pRL-SV40 control vector, followed by doxorubicin (0.3 µg/ml) or mock treatment, according to the schedule described in . Presented are the average-fold luciferase induction relative to FLT1-C mock treated and the standard errors of three replicates. Statistically significant differences relating to the impact of p53 expression alone or in combination with doxorubicin treatment are highlighted (* = p<0.01; ∧ = p<0.05, t-test). (B) The PG13 p53 reporter plasmid was used as a control for comparing the transactivation potential of p53, p73β or p63γ and the effect of the doxorubicin treatment.

Article Snippet: The metastatic neuroblastoma GIMEN cells (p53 wild type) were obtained from GP Tonini (National Institute for Cancer Research, IST, Genoa, Italy), while the colon adenocarcinoma HCT116 (p53 +/+ ) cell line and its p53 −/− derivative and the osteosarcoma-derived SaOS2 cells (p53-null) were a gift from B. Vogelstein (The Johns Hopkins Kimmel Cancer Center, Baltimore, Maryland, USA).

Techniques: Derivative Assay, Transfection, Construct, Over Expression, Plasmid Preparation, Control, Luciferase, Expressing