Thrombospondin-1 Antibody (A6.1) NB100-2059 from Novus Biologicals, a Bio-Techne Brand

Published Figures
Supplier Images

Register or Sign In to View Figures.

eLife
May 11 2022

Fig 1: Tumour conditioned medium (T-cM) induces YAP pathway activation and a foetal-like state in wildtype (WT) organoids.Gene Set Enrichment Analyses (GSEA) showing the correlation between differentially expressed genes in WT organoids cultured in T-cM (A–C) or tumoroids (D–F) and the indicated transcriptional signatures. NES: Normalised Enrichment Score; green NES: positive correlation; red NES: inverse correlation. (G) Representative image of WT organoids expressing Cas9-GFP (in green) transduced with an sgRNA targeting Yap1 (sgYap1 in red). Higher magnification of a budding Yap1KO organoid (in yellow in G') and a cystic Yap1WT organoid expressing only Cas9-GFP but no sgRNA (in green in G''). (H) Percentage of cystic organoids induced by exposure to T-cM in WT, Yap1KO or Tead4KO organoids, as indicated. (I–N) Max projections of immunostaining for YAP1 (in red) of WT organoids exposed to WT-cM (I), or T-cM (J–L) for 24 hr presenting cystic (J) or budding (K, L) morphologies. Organoids in (L) are treated by neutralising antibodies targeting THBS1 (A6.1), which rescues the budding morphology. Organoids in (M) overexpress THBS1 (LentiThbs1) and tumoroids are shown in (N). DAPI stains DNA in blue. White arrowheads pinpoint YAPHIGH cells in Z-section insets. (O) Quantification of the percentage of nuclear YAP (nYAPHIGH) cells/organoid based on the ratio of nuclear vs. cytoplasmic YAP1 in cystic and budding WT organoids grown in WT-cM or T-cM for 24 hr, in WT organoids overexpressing Thbs1 (lenti-Thbs1) or tumoroids, as indicated. (P) Quantification of the percentage of nYAPHIGH cells/organoid in WT organoids cultured with T-cM and control IgG1 or anti-THBS1 (A6.1) antibodies for 24 hr. Scale bars correspond to 100 µm in (G, I–N). Graphs indicate average values ± SD. Statistical analysis was performed with two-tailed unpaired Welch’s t-tests. For the lenti-Thbs1 sample, a Welch’s corrected t-test was applied to compare the percentage of nYAPHIGH cells/organoid between Thbs1-expressing organoids and WT organoids infected with an empty lentivirus. Figure 4—source data 1.Source data related to Figure 4H. Figure 4—source data 2.Source data related to Figure 4O. Figure 4—source data 3.Source data related to Figure 4P.
Register or Sign In to View Figures.

eLife
May 11 2022

Fig 2: Thrombospondin-1 (THBS1) is essential for the growth of tumoroids.(A–D) Representative bright-field images of wildtype (WT) organoids (A, B) or tumoroids (C, D) incubated with IgG1 isotype control antibodies (A, C) or anti-THBS1 A6.1-neutralising antibody (B, D) (10 µg/ml). (E, F) Representative images of tumoroids infected with a lentivirus CRISPR-GFP without sgRNA (control in E) or with an sgRNA targeting Thbs1 (Thbs1-KO in F) 48 hr after replacement of single-cell seeding medium (ENRC) by tumoroid medium (EN). (G, H) Quantification of the number of tumoroids upon antibody neutralisation relative to their size: small tumoroids between 30 and 150 µm in (G); large tumoroids of more than 150 µm diameter in (H). (I) Quantification of the percentage of cystic tumoroids upon treatment by IgG1 isotype control antibodies or three different neutralising antibodies targeting THBS1 (as indicated) for 48 hr. (J) Paired quantification of the number of living tumoroids derived from four independent tumours (from four mice) upon treatment with three neutralising antibodies targeting THBS1 for 48 hr. Antibody concentration: 10 µg/ml. (K–N) Immunofluorescence staining for proliferative cells (EdU in red) and apoptosis (anti-cleaved caspase-3, CASP3 in green) in WT organoids (K, L) or tumoroids (M, N) exposed to IgG1 isotype control antibodies (K, M) or to anti-THBS1 A6.1-neutralising antibody (L, N). (O, P) Quantification of EdU+ cells per organoid (O) or tumoroid (P) in the presence of IgG1 control or anti-THBS1 (A6.1) antibodies. Scale bars = 100 µm. Graphs indicate average values ± SD. Statistical analysis was performed with paired Student’s t-test in (G–J) and two-tailed unpaired Welch’s t-tests in (O) and (P). Figure 3—source data 1.Source data related to Figure 3G. Figure 3—source data 2.Source data related to Figure 3H. Figure 3—source data 3.Source data related to Figure 3I. Figure 3—source data 4.Source data related to Figure 3J. Figure 3—source data 5.Source data related to Figure 3O. Figure 3—source data 6.Source data related to Figure 3P.
Register or Sign In to View Figures.

Technology in cancer research & treatment
January 01 2022

Fig 3: Immunofluorescence staining for Tregs infiltration in high THBS1-expressing samples. (A) Representative image of immunohistochemical staining of THBS1 protein in a high THBS1-expression papillary thyroid carcinoma tissue, scale bar: 100 µm. (B to F) Immunofluorescence staining of Tregs from the same sample (scale bar in B to E: 20 µm, scale bar in F: 10 µm). Arrows point to CD4-positive/FoxP3-positive co-localized Tregs.
Register or Sign In to View Figures.

Technology in cancer research & treatment
January 01 2022

Fig 4: Prognosis analysis of different T-I subgroups and the prognostic value of T-I index in patients with immune therapy. (A) Nomogram of clinical characteristics and T-I index group. (B) Kaplan–Meier survival analysis of T-I index groups. (C) The TIDE, dysfunction and exclusion score in different T-I index groups. (D) ROC curve of T-I index groups and single THBS1 expression groups to predict the responder of immune checkpoint suppression therapy according to the TIDE results. TIDE, the tumor immune dysfunction and exclusion score. ***, P < .001.
Register or Sign In to View Figures.

Technology in cancer research & treatment
January 01 2022

Fig 5: Design of THBS1-immune-related gene prognostic index and K-M plots of 4 selected genes. (A) Venn diagram of 1826 THBS1-related genes and 1793 immune-related genes with 292 genes in intersection region. (B) Co-expression heatmap of THBS1 and other 4 T-I index genes obtained from WGCNA (Spearman analysis, P < .001). (C) Chod diagram of THBS1 and other 4 T-I index genes. The red circle arc represented 2 genes were positively correlated. (D to G) Kaplan–Meier survival analysis of 4 selected genes significant in the univariate Cox analysis (P = .05). T-I index, THBS1-immune-related gene prognostic index; WGCNA, weighted gene co-expression network analysis.