Journal: Clinical and Experimental Immunology

Article Title: Tolerogenic dendritic cells generated with dexamethasone and vitamin D3 regulate rheumatoid arthritis CD4 + T cells partly via transforming growth factor‐ β 1

doi: 10.1111/cei.12870

Figure Lengend Snippet: Transforming growth factor (TGF)‐β1 diminishes the ability of tolerogenic dendritic cells (tolDC) to stimulate healthy control (HC) T cells and is involved in their regulatory function. (a–d) Mature lipopolysaccharide (LPS)‐activated dendritic cells (matDC) or tolDC (1 × 104 cells/well) were co‐cultured with allogeneic HC CD4+ T cells (1 × 105 cells/well) in the absence or presence of increasing concentrations of SB‐505124, a small molecule inhibitor of TGF‐βRI (a) or 1 μM SB‐505124 (b,c) or increasing concentrations of recombinant latency‐associated peptide (LAP) (d). Proliferation (a,b,d, left panels), measured using [3H]‐thymidine uptake, and interferon (IFN)‐γ production (a,b,d, right panels), measured using enzyme‐linked immunosorbent assay (ELISA), were assessed at day 6. (c) Additional cytokines were detected in day 6 culture supernatants from tolDC‐CD4+ T‐cell cultures using an ELISA [interleukin (IL)−17A] or Meso Scale Discovery (MSD) immunoassay (all others). The fold change in cytokine production following inhibition of TGF‐β1 signalling was calculated by: concentration of cytokine produced in the presence of 1μM SB‐505124 ÷ concentration of cytokine produced in the absence of 1 μM SB‐505124. IL‐17A was undetectable in one experiment. Error bars in (a) and (d) represent standard error of the mean (s.e.m.) of triplicates (proliferation) or duplicates (IFN‐γ production). Horizontal lines in (b) and (c) represent median values, (b) left panel (proliferation) n = 7; (b) right panel (IFN‐γ production) n = 15; (c) (cytokines) n = 3. (e) Allogeneic healthy control (HC) CD4+ T cells were primed with DC (10 : 1) for 6 days and rested for 4 days with 10 IU/ml IL‐2. T cell lines primed by matDC (Tmat), tolDC (Ttol) and tolDC + SB‐505124 (Ttol‐SB) were restimulated with matDC and IFN‐γ (left panel) and IL‐10 (right panel) production, measured using ELISA, was assessed on day 3. Results are depicted as the percentage cytokine production of Tmat cell lines. Cytokine concentrations range: IFN‐γ in Ttol = 0.9–20·6 ng/ml and in Ttol‐SB = 4·4–80·7 ng/ml; IL‐10 in Ttol = 0·4–5·1 ng/ml and in Ttol‐SB = 0·9–31·7 ng/ml. Horizontal lines represent median values, n (IFN‐γ production) = 7; n (IL‐10 production) = 6. *P < 0·05 and ***P < 0·0001 calculated with Wilcoxon signed‐rank test. #Significant differences (P < 0·05) between Ttol and Tmat cells.

Article Snippet: TGF‐βRI (ALK5) inhibitor (SB‐505124; Sigma) or recombinant human LAP (R&D Systems) was added where indicated.

Techniques: Control, Cell Culture, Recombinant, Enzyme-linked Immunosorbent Assay, Inhibition, Concentration Assay, Produced

Journal: Anticancer research

Article Title: Intraductal Adaptation of the 4T1 Mouse Model of Breast Cancer Reveals Effects of the Epithelial Microenvironment on Tumor Progression and Metastasis

doi: 10.21873/anticanres.13344

Figure Lengend Snippet: Tumor sphere formation and drug sensitivity. A: Representative image of 4T1 tumor sphere (scale bar=100 μm). B: Numbers of primary and secondary 4T1 tumor spheres (n=3). C: Lapatinib reduced 4T1 tumor sphere formation. D: Inhibitor of activin-like kinase 4/5/7 receptor signaling SB-505124 had no effect on 4T1 tumor sphere formation. Data are presented as the mean±standard error. DMSO: Dimethyl sulfoxide; SFE: Sphere-forming efficiency.

Article Snippet: Lapatinib (Santa Cruz Dallas, TX, USA) and SB-505124 (Sigma) were diluted in dimethyl sulfoxide (DMSO).

Techniques:

Journal: Molecular Oncology

Article Title: LAMC1 upregulation via TGFβ induces inflammatory cancer‐associated fibroblasts in esophageal squamous cell carcinoma via NF‐κB–CXCL1–STAT3

doi: 10.1002/1878-0261.13053

Figure Lengend Snippet: Laminin subunit gamma 1 (LAMC1) expression was upregulated by transforming growth factor β (TGFβ1) via SMAD family member 4 (SMAD4) and SP1 synergistic activation and was associated with a poor prognosis in patients with esophageal squamous cell carcinoma (ESCC). (A) Heatmap of the RNA‐seq data listing genes positively correlated with TGFβ1 in ESCC of GSE53625 ( r > 0.15, P < 0.05). (B) A total of 3084 genes of KYSE30 and KYSE180 were upregulated after TGFβ1 treatment using RNA‐seq data (fold change > 1). (C) Venn diagram of 652 overlapping genes based on positive regulation by TGFβ1 in ESCC. (D) The 31 genes of 652 genes were enriched in 11 pathways involved in the tumor microenvironment. (E,F) LAMC1 was more highly expressed in cancer than in para‐cancer at the RNA and protein levels. Representative immunohistochemical (IHC) images of LAMC1 staining in ESCC tumor tissues and nontumor tissues (original magnification: 200x). (G) Kaplan–Meier survival analysis of OS based on high ( n = 89) and low ( n = 90) LAMC1 expression in GSE53625 (left) and based on high ( n = 35) and low ( n = 20) LAMC1 expression (by IHC, right). (H,I) As verified by western blot and RT‐qPCR, TGFβ1 upregulated LAMC1 expression in a concentration‐ (at different concentrations for 24 h) and time‐dependent manner (at 5 ng·mL −1 for different times) in KYSE30 and KYSE450 cells (H), which could be reversed by the TGFβR1 inhibitor SB505124 (10 μ m ) at the protein (I) and RNA (J) levels. (K,L) After KYSE30 cells were treated with TGFβ1 (5 ng·mL −1 ) for 30 min, the localization of SMAD4 or SP1 to the LAMC1 promoter was detected by ChIP, and the expression of SMAD4 or SP1 was detected in anti‐SP1 or anti‐SMAD4 chromatin fractions separately by western blot (L). (M,N) Knockdown efficiency of SP1 combined with or without SMAD4 knockdown in ESCC cells was verified by western blotting. (O–Q) The expression levels of LAMC1 decreased in ESCC cells with SP1 knockdown combined with (P) or without (O) knockdown, but this effect could not be rescued by TGFβ1 treatment (Q). Three biological replicates were performed for in vitro assays. The data in bar charts are presented as the mean ± SD. ** P < 0.01, *** P < 0.001, **** P < 0.0001 (Student’s t ‐test).

Article Snippet: We used 10 μ m SB505124 and 10 μ m JSH‐23 (Selleck, Houston, TX, USA) to inhibit TGFβ signaling and NF‐κB signaling, respectively.

Techniques: Expressing, Activation Assay, RNA Sequencing, Immunohistochemical staining, Staining, Western Blot, Quantitative RT-PCR, Concentration Assay, Knockdown, In Vitro

Journal: STAR Protocols

Article Title: Protocol for efficient generation of human artery and vein endothelial cells from pluripotent stem cells

doi: 10.1016/j.xpro.2024.103494

Figure Lengend Snippet: Resuspension of lyophilized growth factors and small molecules needed for differentiation

Article Snippet: SB505124 , Tocris , 3263.

Techniques: Concentration Assay

Journal: STAR Protocols

Article Title: Protocol for efficient generation of human artery and vein endothelial cells from pluripotent stem cells

doi: 10.1016/j.xpro.2024.103494

Figure Lengend Snippet: 10 mL of lateral mesoderm differentiation medium (for day 2 of hPSC differentiation)

Article Snippet: SB505124 , Tocris , 3263.

Techniques: Concentration Assay, Generated

Journal: STAR Protocols

Article Title: Protocol for efficient generation of human artery and vein endothelial cells from pluripotent stem cells

doi: 10.1016/j.xpro.2024.103494

Figure Lengend Snippet: 10 mL of vein EC differentiation medium A (for day 3 of hPSC differentiation)

Article Snippet: SB505124 , Tocris , 3263.

Techniques: Concentration Assay, Generated

Journal: STAR Protocols

Article Title: Protocol for efficient generation of human artery and vein endothelial cells from pluripotent stem cells

doi: 10.1016/j.xpro.2024.103494

Figure Lengend Snippet: 10 mL of vein EC differentiation medium B (for day 4 of hPSC differentiation)

Article Snippet: SB505124 , Tocris , 3263.

Techniques: Concentration Assay, Generated

Journal: STAR Protocols

Article Title: Protocol for efficient generation of human artery and vein endothelial cells from pluripotent stem cells

doi: 10.1016/j.xpro.2024.103494

Figure Lengend Snippet:

Article Snippet: SB505124 , Tocris , 3263.

Techniques: Recombinant, Membrane, Pore Size

Journal: Cell Reports Medicine

Article Title: Signaling-induced systematic repression of miRNAs uncovers cancer vulnerabilities and targeted therapy sensitivity

doi: 10.1016/j.xcrm.2023.101200

Figure Lengend Snippet:

Article Snippet: The following drugs were used: 5-Fluorouracil (5-FU), Bleomycin, Cisplatin, Ponatinib, Rapamycin, SB505124 (all Hoelzel Diagnostika); Afatinib, Lapatinib, Linsitinib, Palbociclib (all Biomol); Dabrafenib, Dacomitinib, Pilaralisib, Refametinib, Trametinib (all Selleckchem); GSK690693, Ribociclib (all MedChemExpress); Paclitaxel, SN38 (Biozol).

Techniques: Recombinant, Membrane, Protease Inhibitor, Isolation, Proliferation Assay, Luminescence Assay, Expressing, Sequencing, Comparison, Software

Journal: Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology

Article Title: Targeting cancer cells in the tumor microenvironment: opportunities and challenges in combinatorial nanomedicine

doi: 10.1002/wnan.1358

Figure Lengend Snippet: Nanocarriers for combination drug delivery. The anatomical drawing in the top half of the figure shows three targeting considerations used in nanocarrier design: tissue-, cell-, and macromolecule-level targeting. The bottom half of the figure depicts three novel nanocarriers designed to deliver multiple therapeutic agents to the tumor utilizing one or more of these targeting approaches. Nanocarrier (a) shows a biodegradable nanoscale liposomal polymeric gel engineered to release SB505124, a small molecule inhibitor of the TGFβ1 receptor, as well as IL-2 within the tumor.20 Nanocarrier (b) depicts a particle-within-a-particle encapsulation strategy, whereby dioleoyl phosphatidic acid-coated drug cores were created out of gemcitabine monophosphate (GMP) and cisplatin. GMP and cisplatin drug cores were subsequently loaded into PLGA nanoparticles to generate dual-loaded nanocarriers.21 Of the three nanocarriers depicted here, this nanocarrier is the only one that uses an active (cell-level) targeting approach: Anisamide was attached to the outside of these nanoparticles to target sigma receptor-overexpressing cancer cells. Nanocarrier (c) shows PLGA-PEG/G0-C14 nanoparticles with siRNA molecules contained in their core. A pro-drug form of cisplatin was embedded in the polymeric nanoparticle shell.22

Article Snippet: Nanocarrier (a) shows a biodegradable nanoscale liposomal polymeric gel engineered to release SB505124, a small molecule inhibitor of the TGF β 1 receptor, as well as IL-2 within the tumor.

Techniques: Encapsulation