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: Cancer Medicine

Article Title: A comprehensive analysis of immune infiltration in the tumor microenvironment of osteosarcoma

doi: 10.1002/cam4.4117

Figure Lengend Snippet: The infiltration profile of different immune cells in TME for men and women with osteosarcoma. The analysis was performed using (A) CIBERSORT, (B) MCP‐counter, (C) ssGSEA, and (D) Xcell computational method

Article Snippet: Human osteosarcoma cancer cell lines (HOS, SW1353) were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques:

Journal: Cancer Medicine

Article Title: A comprehensive analysis of immune infiltration in the tumor microenvironment of osteosarcoma

doi: 10.1002/cam4.4117

Figure Lengend Snippet: Consistent clustering diagram of immune cells in different clusters. A heatmap showing the distribution of immune genes (A), and four subgroups of CDF (B). (C) A heatmap showing the infiltration level of immune cells in the four osteosarcoma subgroups. The clusters were generated based on TP, ISs, and SSs

Article Snippet: Human osteosarcoma cancer cell lines (HOS, SW1353) were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Generated

Journal: Cancer Medicine

Article Title: A comprehensive analysis of immune infiltration in the tumor microenvironment of osteosarcoma

doi: 10.1002/cam4.4117

Figure Lengend Snippet: Box plot displaying the expression pattern of immune‐related genes between osteosarcoma subgroups. (A and B) ISs and SSs for the four osteosarcoma clusters. Compared with the other three clusters, cluster 4 had a higher immune score. (C–F) The expression levels of multiple immune genes among the four clusters. *, p < 0.05. **, p < 0.01. ***, p < 0.001. ****, p < 0.0001

Article Snippet: Human osteosarcoma cancer cell lines (HOS, SW1353) were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Expressing

Journal: Cancer Medicine

Article Title: A comprehensive analysis of immune infiltration in the tumor microenvironment of osteosarcoma

doi: 10.1002/cam4.4117

Figure Lengend Snippet: Distribution of the main pathological osteosarcoma subtypes in the four osteosarcoma clusters. (A–D) The major pathological subtypes including Leiomyosarcoma (LMS), Myxofibrosarcoma (MF), undifferentiated pleomorphic sarcoma (MFH), and Undifferentiated Pleomorphic Sarcoma (UPS) in the four clusters. (E and F) The survival rate of patients in different subtypes

Article Snippet: Human osteosarcoma cancer cell lines (HOS, SW1353) were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques:

Journal: Cancer Medicine

Article Title: A comprehensive analysis of immune infiltration in the tumor microenvironment of osteosarcoma

doi: 10.1002/cam4.4117

Figure Lengend Snippet: Validation of NPTX2 function in osteosarcoma. (A–C) Association of NPTX2 mRNA with CD8A, IFNG, and GZMB expression in osteosarcoma tissues ( n = 35). (D) The knockdown efficacy of NPTX2 in HOS and SW1353 cells as determined by qPCR. (E and F) Cell proliferation was measured with the CCK8 assays in HOS and SW1353 cells transfected with siRNAs or NC. (G and H) Apoptosis of HOS cells and SW1353 cells after transfection with siRNAs or NC was detected by flow cytometric assay. (I and J) The expression of CCL4, CCL5, CXCL9, CXCL10, and CXCL13 as measured with qPCR in HOS‐NPTX2‐NC and HOS‐NPTX2‐siRNAs cells, or SW1353‐NPTX2‐NC and SW1353‐NPTX2‐siRNAs cells. (K and L) The expression of CCL4, and CXCL13 as quantified with ELISA assay. (M and N) Association of NPTX2 mRNA with CCL4 expression in the TCGA datasets and in osteosarcoma tissues from our patients ( n = 35). (O and P) Association of CCL4 mRNA with CD8A expression in the TCGA datasets and osteosarcoma tissues from our patients ( n = 35)

Article Snippet: Human osteosarcoma cancer cell lines (HOS, SW1353) were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Biomarker Discovery, Expressing, Knockdown, Transfection, Flow Cytometry, Enzyme-linked Immunosorbent Assay

Journal: Cancer Medicine

Article Title: A comprehensive analysis of immune infiltration in the tumor microenvironment of osteosarcoma

doi: 10.1002/cam4.4117

Figure Lengend Snippet: Hub genes for the osteosarcoma prognostic model. (A) Multivariate cox regression analysis of hub genes. (B) Kaplan–Meier curves showing the OS of patients with high and low prognostic scores. (C) The time‐dependent ROC curves displaying the prognostic prediction of the hub genes model

Article Snippet: Human osteosarcoma cancer cell lines (HOS, SW1353) were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques:

Journal: Cancer Medicine

Article Title: A comprehensive analysis of immune infiltration in the tumor microenvironment of osteosarcoma

doi: 10.1002/cam4.4117

Figure Lengend Snippet: Relationship between clinical features and prognosis of osteosarcoma. (A) Multivariate cox regression analysis results indicating the relationship between metastasis, risk score, and OS. (B) The ROC showing the prognostic accuracy of the 11‐gene model across several cancer and disease factors (gender, across age, in metastatic, and non‐metastatic tumors as well as depth, length, and width). (C) The predictive accuracy of the 11‐gene risk score model in predicting immunotherapy response. The low‐risk group had better OS than the high‐risk group ( p < 0.0001)

Article Snippet: Human osteosarcoma cancer cell lines (HOS, SW1353) were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques:

Journal: Nature Communications

Article Title: Direct single-molecule detection and super-resolution imaging with a low-cost portable smartphone-based microscope

doi: 10.1038/s41467-025-63993-z

Figure Lengend Snippet: a Sketch of an immunolabeled microtubule with primary and secondary antibodies. The secondary antibody is conjugated with a docking DNA strand complementary to the Cy3B-labeled imager strand. b Super-resolution and diffraction-limited images of the microtubule network of fixed U2OS cells obtained with the smartphone-based microscope. The super-resolved image represents the localization density, while the diffraction-limited image is the actual fluorescence intensity. Scale bar: 2 µm. c , d Comparison of super-resolved images of the microtubule network obtained with the smartphone-based setup ( c ) – a magnified image of the region marked in ( b ) - and with the high-end microscope ( d ). Scale bar: 2 µm. e Line profiles across super-resolved microtubules as indicated in ( c ) and ( d ). Black solid lines: smartphone-based microscope. Pink dashed lines: high-end microscope. In total, four different videos were obtained in which the immunolabeled microtubules could be resolved with the smartphone-based microscope. Source data are provided as a Source Data file.

Article Snippet: A human bone osteosarcoma epithelial (U2OS) cell culture with immunolabeled microtubules was purchased from Massive Photonics.

Techniques: Immunolabeling, Labeling, Microscopy, Fluorescence, Comparison