Journal: Theranostics

Article Title: An αvβ6/αvβ8-targeting peptibody inhibits integrin-dependent TGFβ activation, enables tumor-selective cytotoxic delivery, and synergizes with PD-L1 immune checkpoint blockade

doi: 10.7150/thno.131237

Figure Lengend Snippet: 4ΔmFc and peptide 5a, but not mIgG1 and peptide 2a (negative controls) inhibit TGFβ-activation mediated by 5M701/GFP/CEA and 5637 cancer cells. Cells were plated in 96-well microtiter plates, left to adhere (2 h), and exposed to the indicated proteins or peptides for 24-48 h. The concentration of active TGFβ in the supernatants was assessed using a bioassay based on HEK-Blue™ TGFβ cells, as detailed in the Methods . A representative experiment out of two performed is shown (mean ± SE, n = 3-4 wells). The EC 50 of each compound, calculated from the results of both experiments, is shown.

Article Snippet: Human TGFβ1 was from InvivoGen (cat. rcyc-htgfb1).

Techniques: Activation Assay, Concentration Assay, Bioassay

Journal: Theranostics

Article Title: An αvβ6/αvβ8-targeting peptibody inhibits integrin-dependent TGFβ activation, enables tumor-selective cytotoxic delivery, and synergizes with PD-L1 immune checkpoint blockade

doi: 10.7150/thno.131237

Figure Lengend Snippet: 4ΔmFc reduces active TGFβ levels and enhances immune cell infiltration in the TS/A tumor microenvironment. A ) Experimental scheme. TS/A mammary adenocarcinoma cells were implanted subcutaneously in 35 mice. On day 6 after tumor implantation, tumor-bearing mice were randomized into two experimental groups and treated intraperitoneally with vehicle alone (Group G1: n = 17 mice) or with 50 µg of 4Δ mFc (Group G2: n = 18 mice), at the times indicated by the arrows. Tumors were collected on day 13, weighed, and randomly assigned to either TGFβ quantification by ELISA (n = 9 tumors/group) or flow cytometry analysis (n = 8–9 tumors/group). B ) Tumor weight. Bars, mean ± SE of n = 17-18 tumors/group. Dots represent individual tumors. P value, calculated by two-tailed unpaired t-test, is indicated in the plot. C ) Active and total TGFβ levels in TS/A tumor homogenates as determined by ELISA. Bars, mean ± SE of n = 9 tumors/group. Dots represent individual tumors. P value was calculated by two-tailed unpaired t-test and is indicated in the plot. D ) Flow cytometry quantification of tumor-infiltrating immune cells, expressed as cells per mg of tumor. Dots represent individual tumors; box plots indicate the median and interquartile range (n = 8-9 tumors/group). P values were calculated by two-tailed unpaired t-test and are indicated in the plots.

Article Snippet: Human TGFβ1 was from InvivoGen (cat. rcyc-htgfb1).

Techniques: Tumor Implantation, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Two Tailed Test

Journal: Cell Reports Medicine

Article Title: Human astrocytes and microglia show augmented ingestion of synapses in Alzheimer’s disease via MFG-E8

doi: 10.1016/j.xcrm.2023.101175

Figure Lengend Snippet:

Article Snippet: Recombinant human TGFβ-1 , Miltenyi , 170-076-166.

Techniques: Labeling, Blocking Assay, Cell Isolation, Recombinant, Plasmid Preparation, Protease Inhibitor, Western Blot, Molecular Weight, Marker, Software

Journal: Frontiers in Pharmacology

Article Title: FGFR1/YAP1 signaling in endothelial cells drives renal fibrosis and offers a therapeutic target

doi: 10.3389/fphar.2026.1766265

Figure Lengend Snippet: TGF-β induces endothelial cell profibrotic activation via the FGFR1/YAP1 axis. (A) Flow cytometry verifying >99% purity of isolated human umbilical vein endothelial cells (HUVECs) by CD31 expression. (B) Morphological changes of HUVECs treated with TGF-β (2.5, 7.5, 22.5 ng/ml): concentration-dependent transition from cobblestone to spindle-shaped, smooth muscle cell-like phenotype. Bar = 20 μm. (C) qPCR analysis of fibrotic markers (ACTA2, Vimentin) and YAP1 mRNA in TGF-β-stimulated HUVECs. Data are fold change vs. unstimulated control (set as 1). (D) Western blot showing TGF-β-induced upregulation of α-SMA, FGFR1, and YAP1 in HUVECs after 24-h treatment (0, 2.5, 7.5, 22.5 ng/ml). Data are fold change vs. unstimulated control (set as 1). (E–G) Semi-quantitative Western blot confirming TGF-β-induced upregulation of α-SMA (dose-dependent; (E) , FGFR1 (F) , and YAP1 (G) in HUVECs. n = 3. Data are presented as mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, no significance.

Article Snippet: To establish an in vitro fibrosis model, cells were first synchronized by serum-free starvation for 12 h and then stimulated for 24 h with recombinant human TGF-β (Novoprotein Biotech, #CA59; 0, 2.5, 7.5, or 22.5 ng/ml; reconstituted in distilled water).

Techniques: Activation Assay, Flow Cytometry, Isolation, Expressing, Concentration Assay, Control, Western Blot

Journal: Frontiers in Pharmacology

Article Title: FGFR1/YAP1 signaling in endothelial cells drives renal fibrosis and offers a therapeutic target

doi: 10.3389/fphar.2026.1766265

Figure Lengend Snippet: PD173074 , an FGFR1 inhibitor, mitigates TGF-β-induced EndMT via targeting the FGFR1/YAP1 axis (A) Experimental protocol: HUVECs were seeded in 35 mm dishes, adhered for 12 h, then treated with 22.5 ng/ml TGF-β for 24 h to induce fibrosis, followed by intervention with 22.5 ng/ml TGF-β plus PD173074 (0, 10, 30, 90 nM). (B) Phase-contrast images showing PD173074 (30, 90 nM) reversed TGF-β-induced morphological transition from spindle-shaped to cobblestone-like phenotype. Bar = 20 μm (C) qPCR analysis of FGFR1, YAP1, and EndMT markers (ACTA2, Vimentin) mRNA in HUVECs under indicated conditions. Data normalized to GAPDH, presented as fold change vs. untreated control (set to 1). n = 3. (D) Western blot confirming PD173074 -induced downregulation of FGFR1, YAP1, and α-SMA, consistent with GAPDH-normalized semi-quantitative data (fold change vs. untreated control, set to 1). n = 4. Data are presented as mean +SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, no significance.

Article Snippet: To establish an in vitro fibrosis model, cells were first synchronized by serum-free starvation for 12 h and then stimulated for 24 h with recombinant human TGF-β (Novoprotein Biotech, #CA59; 0, 2.5, 7.5, or 22.5 ng/ml; reconstituted in distilled water).

Techniques: Control, Western Blot

Journal: Frontiers in Pharmacology

Article Title: FGFR1/YAP1 signaling in endothelial cells drives renal fibrosis and offers a therapeutic target

doi: 10.3389/fphar.2026.1766265

Figure Lengend Snippet: FGFR1 or YAP1 knockdown attenuates EndMT (A,B) Validation of FGFR1 knockdown efficiency in HUVECs at the protein (A) and mRNA (B) levels. (C) Western blot analysis demonstrated that TGF-β induces the upregulation of α-SMA, YAP1, and FGFR1 expression in shNC cells, while knockdown of FGFR1 inhibits this effect. (D) Morphological reversion of HUVECs from spindle-shaped (TGF-β-induced) to cobblestone-like after FGFR1 knockdown. Bar = 20 μm. (E) qPCR analysis showing TGF-β-induced upregulation of ACTA2, YAP1, ACTA2 and Vimentin in control cells, which was suppressed by FGFR1 knockdown. (F,G) Validation of YAP1 knockdown efficiency at the protein (F) and mRNA (G) levels. (H) qPCR analysis of ACTA2 and Vimentin mRNA confirming TGF-β-induced fibrosis and suppression by YAP1 knockdown. (I) Western blot analysis demonstrated that YAP1 knockdown significantly suppressed the TGF-β-induced upregulation of α-SMA and YAP1 expression. (J) Morphological reversion of HUVECs to cobblestone-like after YAP1 knockdown.​ Bar = 20 μm. All data are presented as mean +SD (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, no significance. mRNA data were normalized to GAPDH or β-actin and expressed as fold change vs. scrambled control (set to 1).

Article Snippet: To establish an in vitro fibrosis model, cells were first synchronized by serum-free starvation for 12 h and then stimulated for 24 h with recombinant human TGF-β (Novoprotein Biotech, #CA59; 0, 2.5, 7.5, or 22.5 ng/ml; reconstituted in distilled water).

Techniques: Knockdown, Biomarker Discovery, Western Blot, Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: FGFR1/YAP1 signaling in endothelial cells drives renal fibrosis and offers a therapeutic target

doi: 10.3389/fphar.2026.1766265

Figure Lengend Snippet: FGFR1 knockdown attenuates inflammation and endothelial dysfunction and reveals underlying mechanisms (A) qPCR analysis showed that FGFR1 knockdown reduced the mRNA levels of VCAM1, IL-6, and TNF-α under TGF-β stimulation. (B) Western blot analysis confirmed that FGFR1 knockdown decreased the protein expression of VCAM1 and IL-6 under TGF-β stimulation. (C) Under TGF-β stimulation, FGFR1 knockdown downregulated Ang2 mRNA expression compared with the shNC group. (D) FGFR1 knockdown decreased p-VE-cadherin protein expression under TGF-β stimulation relative to the shNC group. (E) Under TGF-β stimulation, FGFR1 knockdown reduced CCN2 mRNA expression compared with the shNC group. (F) Western blot analysis revealed that FGFR1 knockdown downregulated p-ERK and YAP1 protein levels under TGF-β stimulation relative to the shNC group. All data are presented as mean +SD (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, no significance. mRNA data were normalized to GAPDH and expressed as fold change vs. scrambled control (set to 1).

Article Snippet: To establish an in vitro fibrosis model, cells were first synchronized by serum-free starvation for 12 h and then stimulated for 24 h with recombinant human TGF-β (Novoprotein Biotech, #CA59; 0, 2.5, 7.5, or 22.5 ng/ml; reconstituted in distilled water).

Techniques: Knockdown, Western Blot, Expressing, Control