Journal: Life Science Alliance

Article Title: Variants of Epas1 contribute to hypoxia adaptation in the subterranean rodents Eospalax and Spalax

doi: 10.26508/lsa.202603622

Figure Lengend Snippet: (A) mRNA levels of Epas1 and its target gene Cited2 were significantly higher in E. baileyi LFs compared with rats under normoxia, whereas Col3a1 was much lower in E. baileyi LFs. (B) Hypoxia up-regulated the mRNA expression of Col3a1 in rat LFs, but only induced Epas1 gene expression in E. baileyi LFs. (C) TGF-β treatment significantly induced an increase in Col3a1 mRNA levels in rat LFs, but in E. baileyi LFs, Epas1 and Cited2 mRNA levels significantly increased, whereas Col3a1 mRNA showed no significant change. (D) Western blot analysis revealed that hypoxia promoted COL1A1 protein expression in rat cells, whereas hypoxia inhibited COL1A1 protein expression in E. baileyi LFs. (E) mRNA expressions of collagen family genes and interferon-related genes were decreased in E. baileyi LFs under hypoxia. (F) COL1A1 protein showed much lower fluorescence intensity in E. baileyi LFs compared with rat cells in immunofluorescence analysis. (G, H) TGF-β treatment significantly induced fibrosis in primary rat LFs, but had no significant effect on E. baileyi LFs. Scale bar: 30 μm. Data are the mean ± SEM, with each point representing one replicate mean. Statistical comparisons among different treatments were performed using a one-way ANOVA and paired t test. * P < 0.05, ** P < 0.01, *** P < 0.001 versus rat LF or control, n = 3.

Article Snippet: Cells were moved to the hypoxia incubator in which the O 2 level was 0.1% for 24 h. Cells were treated with 10 μM Raf kinase inhibitor LY3009120, 10 μM ATM kinase inhibitor KU-55933, 20 μM PKA inhibitor H89 2HCL, 10 μg/ml specific inhibitor of AKT MK-2206 2HCL (Selleck), or 10 μM EPAS1 inhibitor belzutifan PT2977 (MCE) for 24 h, 5 ng/ml TGF-β (MCE) for 48 h, and 1 M H 2 O 2 for 6 h before qRT-PCR, Western blot, and immunofluorescence analyses.

Techniques: Expressing, Gene Expression, Western Blot, Fluorescence, Immunofluorescence, Control

Journal: bioRxiv

Article Title: Fitness and immune-escape within germinal centers shape premalignant evolution toward lymphoma

doi: 10.64898/2026.06.25.734549

Figure Lengend Snippet: (A) Experimental design for single-cell RNA sequencing of PD-1⁺ CD8⁺ T-cells sorted from recipient mice 10 days after immunization. (B) UMAP projection of PD-1⁺ CD8⁺ T-cells colored by annotated subset identity. (C) Dot plot showing expression of selected marker genes across annotated CD8⁺ T-cell subsets. (D) Density plots showing the distribution of PD-1⁺CD8⁺ T-cells from Ctrl, BC, and BCK recipients across the UMAP. (E) UMAP projections showing expression of selected marker genes from (C). (F) Slingshot trajectory analysis of PD-1⁺ CD8⁺ T-cell differentiation, highlighting trajectory path1 from activated states toward stem-like, early effector and effector states. (G) Comparison of CD8⁺ T-cell subset distribution along path 1, which terminates in cytotoxic effector differentiation. Quantification shows mean cluster proportion from two biological replicates. (H) Gene set enrichment analysis of HALLMARK pathways comparing cluster 4 from BCK and BC conditions. (I-K) TGF-β receptor blockade following GC response derived from transferred BCK B-cells. (I) Experimental design for LY2109761 treatment. (J) Left, representative gating of granzymeB⁺ perforin⁺ effector cells among PD-1⁺ CD8⁺ T-cells. Right, quantification of PD-1⁺ CD8⁺ T-cells and granzymeB⁺ perforin⁺ effector CD8⁺ T-cells per 10⁶ splenocytes. (K) Left, representative gating of donor-derived reporter⁺ CD45.2⁺ B-cells. Right, quantification of donor-derived reporter⁺ CD45.2⁺ GC B-cells per 10⁶ splenocytes. (L) Endogenous CD45.1⁺ GC B-cell response to SRBCs in the absence or presence of TGF-β receptor inhibition with LY2109761. Left representative gating of host CD45.1⁺ GC B-cell B-cells. Right, quantification of host CD45.1⁺ GC B-cell B-cells. (M) Requirement for CD8⁺ T-cells during TGF-β receptor blockade. Top, schematic of experimental design. Bottom, representative gating of donor-derived reporter⁺ CD45.2⁺ B-cells from BCK recipients treated with LY2109761 together with isotype control or anti-CD8 antibody; and quantification of reporter⁺ CD45.2⁺ GC B-cells per 10⁶ splenocytes. Each symbol in (J, L, K, M) represents an individual mouse. Each symbol in (G) represents mean value from two biological replicates. (J-L) placebo n = 3, LY2109761 n = 3. (M) LY2109761 + isotype n = 3, LY2109761 + anti-CD8 n = 3. Small horizontal lines indicate mean ± SD. Data in (J-L) are representative of two independent experiments; data in (K) are representative of two independent experiments. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001; Two-way ANOVA in (G) , value calculated on two biological replicates from each group, remaining statistical calculations used unpaired two-tailed Student’s t test. ns, not significant.

Article Snippet: For TGF-β pathway inhibition, mice were treated with the dual TGF-β type I/II receptor inhibitor LY2109761 (MedChemExpress, HY-12075; 40 mg/kg).

Techniques: Single Cell, RNA Sequencing, Expressing, Marker, Cell Differentiation, Comparison, Derivative Assay, Inhibition, Control, Two Tailed Test

Journal: Biomedicines

Article Title: THBS1 Induces Dysfunction of Ovarian Granulosa Cells in Patients with Polycystic Ovary Syndrome by Activating the TGF-β/Smad Pathway

doi: 10.3390/biomedicines14061273

Figure Lengend Snippet: THBS1 overexpression activates the TGF-β1/Smad axis. ( A ) DESeq2 volcano (267 up + 317 down at |log FC| > 1, Padj < 0.05). ( B ) Hierarchical clustering of differentially expressed genes. ( C ) TGF-β/Smad pathway gene expression (28 canonical genes). *, **, *** Data are presented as mean ± SEM. ( D ) Reciprocal co-immunoprecipitation: lanes IgG/IP/Input. ( E ) Active TGF-β1—patient FF. Different colored bars represent different experimental groups, and different symbols indicate individual sample data points. ( F ) Active/Total ratio--patient FF. ( G ) Active TGF-β1—rat serum. ( H ) Active/Total ratio—rat serum. ( I ) Western blot analysis of TGF-β/Smad—KGN cells (THBS1-OE). ( J , K ) Flow cytometry apoptosis (THBS1-OE ± SB-431542). ( L – N ) Western blot analysis of apoptosis markers (BAX, BCL-2), inflammatory cytokines (IL-6, TNF-α), steroidogenic enzymes (CYP19, CYP17), and TGF-β/SMAD2 signaling pathway components.

Article Snippet: Finally, to determine whether the cellular dysfunction induced by THBS1 overexpression depends on the TGF-β receptor, we treated THBS1-OE KGN cells with the type-I receptor (ALK4/5/7) inhibitor SB-431542 (MedChemExpress HY-10431, 10 μM, 48 h).

Techniques: Over Expression, Gene Expression, Immunoprecipitation, Western Blot, Flow Cytometry

Journal: bioRxiv

Article Title: Fasting disrupts the InsP₆–HDAC3 axis to drive ER stress–mediated clearance of DNA-damaged cells and enforce tissue quality control

doi: 10.64898/2026.05.24.727426

Figure Lengend Snippet: (A) HDAC3 activity assay in GFD-treated HCT116 cells following stimulation (3 h) with TGFβ, IGF1, VEGF, EGF, FGF, or PDGF. (n = 3; mean ± SD). (B) Immunoblot analysis of H3K9ac, H3K27ac, H4K16ac, and HDAC3 in untreated, GFD, and growth factor-stimulated conditions. (n = 3). (C) Co-immunoprecipitation of HDAC3 from HCT116 cells overexpressing FLAG-SMRT deacetylase-activating domain (FLAG-DAD) under indicated conditions. (n = 3). (D) Immunoblot analysis of IPMK, CDK5RAP3, CHOP, IRE1, and H3K27ac following treatment with the TGFβ receptor inhibitor vactosertib in HCT116 cells. (n = 3). (E) HDAC3 enzymatic activity assay in HCT116 cells treated with TGF-β, and vactosertib. (n = 3; mean ± SD). (F) Immunoblot analysis of H3K27ac in untreated, GFD, vactosertib-treated, and HDAC3 knockout cells. (n = 3). (G) Immunoprecipitation of IPMK–myc followed by immunoblotting for FBXO22, SKP1, Cullin1, and K48-linked ubiquitin under the indicated conditions. (n = 3). (H) Co-immunoprecipitation of phosphorylated SMAD2/3 with FBXO22, SKP1, and Cullin1 under the indicated conditions. (n = 3). (I) Co-immunoprecipitation of phosphorylated SMAD2/3 with FBXO22–SCF–Cullin1 components following vactosertib treatment. (n = 3). (J) IPMK expression and pSMAD2/3 level after Verteportin (blocks pSmad2/3 nuclear localization) treatment. (n = 3). (K) Immunoprecipitation study to show effects of Verteportin on FBXO22 interaction with IPMK and pSMAD2/3. (n = 3).

Article Snippet: For the inhibition of TGF-β signaling, cells were treated with 1 μM Vactosertib (Cat. No. 19928 MCE chemicals) or 50 μM RepSoX (Cat no. HY-13012 MCE Chemical) overnight (approximately 16 hours) in complete growth media supplemented with serum.For the enteroid system, RepSOX treatment was administered for 6 hr at a dose of 50 μM.

Techniques: Activity Assay, Western Blot, Immunoprecipitation, Histone Deacetylase Assay, Enzyme Activity Assay, Knock-Out, Ubiquitin Proteomics, Expressing

Journal: Frontiers in Immunology

Article Title: Hepatitis C virus core protein-induced myeloid-derived suppressor cells promote hepatic fibrosis by regulating hepatic stellate cell function via TGF-β

doi: 10.3389/fimmu.2026.1795273

Figure Lengend Snippet: Comparison of peripheral blood CD14+HLA-DR-/low monocyte proportion and TGF-β levels between CHC and HC groups. (A) CD14 + monocytes were isolated and purified from CHC patients and healthy donors using magnetic bead sorting technology. Specific antibody staining followed by flow cytometric analysis determined the proportion of CD14+HLA-DR-/low phenotype MDSCs. (B) Statistical analysis of CD14 + HLA-DR-/ low cells in 10 healthy donors and 10 patients with chronic hepatitis C. (C) ELISA detection of TGF-β expression differences in serum between the CHC and HC groups. Each experiment was performed with samples from ten independent donors (n = 10). Error bars represent standard errors of the mean. ***P < 0.001.

Article Snippet: TGF-β inhibitor was purchased from MedChemExpress (New Jersey, USA).

Techniques: Comparison, Isolation, Purification, Staining, Enzyme-linked Immunosorbent Assay, Expressing

Journal: Frontiers in Immunology

Article Title: Hepatitis C virus core protein-induced myeloid-derived suppressor cells promote hepatic fibrosis by regulating hepatic stellate cell function via TGF-β

doi: 10.3389/fimmu.2026.1795273

Figure Lengend Snippet: MDSCs regulate LX2 proliferation, apoptosis and activation via TGF-β signaling. As shown, TGF-β expression was markedly higher in the MDSC group than in the CD14 + monocyte group. MDSCs were co-cultured with LX2 cells for three days with or without TGF-β inhibitor. (A) ELISA detection of TGF-β differences between MDSC and CD14 + monocyte groups. (B) Regulation of LX2 by MDSCs at different concentrations of CAT-192. (C) MTT assay for LX2 proliferation. (D) ELISA detection of LX2 Col-1 secretion. (E) Flow cytometry analysis of LX2 apoptosis. (F) Representative flow cytometric analysis of LX2 apoptosis. Each condition was tested in three independent experiments (n = 3). Error bars denote standard error of the mean. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001.

Article Snippet: TGF-β inhibitor was purchased from MedChemExpress (New Jersey, USA).

Techniques: Activation Assay, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, MTT Assay, Flow Cytometry

Journal: Frontiers in Immunology

Article Title: Hepatitis C virus core protein-induced myeloid-derived suppressor cells promote hepatic fibrosis by regulating hepatic stellate cell function via TGF-β

doi: 10.3389/fimmu.2026.1795273

Figure Lengend Snippet: Schematic diagram of the mechanism by which hepatitis C virus core protein-induced myeloid-derived suppressor cells promote hepatic fibrosis by regulating hepatic stellate cell function via TGF-β.

Article Snippet: TGF-β inhibitor was purchased from MedChemExpress (New Jersey, USA).

Techniques: Virus, Derivative Assay, Cell Function Assay

Journal: Frontiers in Pharmacology

Article Title: Dasatinib and quercetin mitigate radiation-induced lung injury by eliminating senescent cells in a rat model

doi: 10.3389/fphar.2026.1748788

Figure Lengend Snippet: DQ suppresses radiation-induced SASP activation in lung tissue. (A) Representative Western blot images and quantitative analysis of IL-6, IL-1β, IL-18, TNF-α, TGF-β, α-SMA, MMP2, MMP9 (n = 3). (B) Representative immunohistochemical staining images (magnification ×200) and quantitative analysis of TGF-β and MCP1 expression in lung tissues (n = 3). Data are presented as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Sections were incubated overnight at 4 °C with the indicated primary antibodies against TGF-β (YP-Ab-15967,1:200, UpingBio, China) and MCP-1 (HY- P81171 ,1:200, MCE, China), followed by incubation with HRP-conjugated secondary antibodies.

Techniques: Activation Assay, Western Blot, Immunohistochemical staining, Staining, Expressing