Journal: bioRxiv

Article Title: TGFβ-induced long non-coding RNA LINC00313 activates Wnt signalling and promotes cholangiocarcinoma

doi: 10.1101/2022.09.28.509889

Figure Lengend Snippet: TGFβ-mediated LINC00313 up-regulation is CCA-specific. A) Real-time qPCR for detection of LINC00313 specific exonic regions in HuCCT1 or Huh28 cells treated with the TβRI inhibitor LY2157299 or DMSO, and stimulated with TGFβ1 or BSA/HCl for 16h. B) LINC00313 expression levels (normal logarithmic scale) among every cancer type of the PanCancer Atlas of TCGA, sorted from the highest (top) to the lowest (bottom) expression and real-time qPCR to detect LINC00313 expression in HCC cell lines HepG2 and Hep3B treated as in panel A.

Article Snippet: The TβRI inhibitor LY2157299 (Interchim, LSK040) was added to the cells at a final concentration of 10 µM.

Techniques: Expressing

Journal: bioRxiv

Article Title: TGFβ-induced long non-coding RNA LINC00313 activates Wnt signalling and promotes cholangiocarcinoma

doi: 10.1101/2022.09.28.509889

Figure Lengend Snippet: A) LINC00313 expression in HuCCT1 and Huh28 cells treated with LY2157299 or DMSO, and stimulated with TGFβ1 or BSA/HCl. B) LINC00313 and SERPINE1 expression in HuCCT1 and Huh28 treated with SIS3 or DMSO and stimulated with TGFβ1 or BSA/HCl. C) LINC00313 expression in HuCCT1 transiently transfected with siRNA targeting SMAD2 , SMAD3 , or SMAD4 , alone or in combination, with or without TGFβ1. D) LINC00313 expression in Huh28 transiently transfected with siRNA targeting SMAD2 , SMAD3 , or SMAD4 , alone or in combination, with or without TGFβ1. E) LINC00313 levels in HuCCT1 or Huh28 treated with MEK, p38, JNK or PI3K inhibitors, with or without TGFβ1 stimulation.

Article Snippet: The TβRI inhibitor LY2157299 (Interchim, LSK040) was added to the cells at a final concentration of 10 µM.

Techniques: Expressing, Transfection

Journal: bioRxiv

Article Title: TGFβ-induced long non-coding RNA LINC00313 activates Wnt signalling and promotes cholangiocarcinoma

doi: 10.1101/2022.09.28.509889

Figure Lengend Snippet: LINC00313 is induced by TGFβRI-SMAD signalling. A) Real-time qPCR for detection of LINC00313 in eCCA cell line Sk-ChA-1 and in iCCA cell lines CCLP1, SG231 treated with the TβRI inhibitor LY2157299 or DMSO, and stimulated with TGFβ1 or BSA/HCl for 16h. B) Real-time qPCR to measure SMAD2 , SMAD3 and SMAD4 mRNA levels in HuCCT1 and Huh28 cell lines transiently transfected with siRNAs targeting SMAD2 , SMAD3 or SMAD4 or a non-targeting siRNA (siNC) and stimulated with TGFβ1 or BSA/HCl for 16h.

Article Snippet: The TβRI inhibitor LY2157299 (Interchim, LSK040) was added to the cells at a final concentration of 10 µM.

Techniques: Transfection

Journal: Heliyon

Article Title: Macrophage expression of constitutively active TβRI alleviates hepatic injury in a mouse model of concanavalin A-induced autoimmune hepatitis

doi: 10.1016/j.heliyon.2025.e42691

Figure Lengend Snippet: Nucleotide sequences of qPCR primers.

Article Snippet: TβRI CA mice were kindly provided by Professor Laurent Bartholin, INSERM, France.

Techniques:

Journal: Heliyon

Article Title: Macrophage expression of constitutively active TβRI alleviates hepatic injury in a mouse model of concanavalin A-induced autoimmune hepatitis

doi: 10.1016/j.heliyon.2025.e42691

Figure Lengend Snippet: Expression analysis of TβRI CA transgenes in livers of TβRI CA /Fsp1-Cre mice. (A) Detection of TβRI CA transcripts by qPCR performed on total RNA from livers of wild-type and TβRI CA /Fsp1-Cre mice. (B) Expression of Tgf-β, Snail1, Twist and PAI-1 in livers of wild-type and TβRI CA /Fsp1-Cre mice (n = 6). (C) Phosphorylated form of Smad2 and 3 (pSmad2/3) level was analyzed by Western blot analysis. β-actin proteins served as a control. Non-adjusted images of pSmad2/3 and β-actin Western blots can be found in supplemental materials. (D) Hematoxylin/Eosin and Sirius Red/Fast green staining of liver sections obtained from control and TβRI CA /Fsp1-Cre mice. The mRNA expression of TβRI CA (E) and Snail1 (F) in macrophages isolated from livers of wild-type and TβRI CA /Fsp1-Cre mice (n = 6). Data are shown as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗∗ P < 0.0001 between the two indicated groups.

Article Snippet: TβRI CA mice were kindly provided by Professor Laurent Bartholin, INSERM, France.

Techniques: Expressing, Western Blot, Control, Staining, Isolation

Journal: Heliyon

Article Title: Macrophage expression of constitutively active TβRI alleviates hepatic injury in a mouse model of concanavalin A-induced autoimmune hepatitis

doi: 10.1016/j.heliyon.2025.e42691

Figure Lengend Snippet: Fsp1-Cre- mediated expression of TβRI CA is protective against conA-induced liver injury. (A–B) Serum level of aspartate transaminase (AST) and alanine transaminase (ALT) in control and TβRI CA /Fsp1-Cre mice treated with PBS or conA for 6 h. AST (A) and ALT (B) level was lower in TβRI CA /Fsp1-Cre mice compared to those of control mice. Data are means ± SD of n = 10 per experimental group. (C) Survival experiments were performed in wild-type and TβRI CA /Fsp1-Cre mice treated with conA 20 mg/kg body weight (n = 11–12 per group). ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001 between the two indicated groups.

Article Snippet: TβRI CA mice were kindly provided by Professor Laurent Bartholin, INSERM, France.

Techniques: Expressing, Control

Journal: Heliyon

Article Title: Macrophage expression of constitutively active TβRI alleviates hepatic injury in a mouse model of concanavalin A-induced autoimmune hepatitis

doi: 10.1016/j.heliyon.2025.e42691

Figure Lengend Snippet: Assessment of liver histopathology following conA treatment. (A) Paraffin sections of liver from conA-treated wild-type and TβRI CA /Fsp1-Cre mice. The fraction of necrotic area was enclosed by dash lines and percentage necrotic area was calculated by examining 10 high-power fields/livers (B). Data are shown as means ± SD. ∗∗∗∗ P < 0.0001 between the two indicated groups.

Article Snippet: TβRI CA mice were kindly provided by Professor Laurent Bartholin, INSERM, France.

Techniques: Histopathology

Journal: Heliyon

Article Title: Macrophage expression of constitutively active TβRI alleviates hepatic injury in a mouse model of concanavalin A-induced autoimmune hepatitis

doi: 10.1016/j.heliyon.2025.e42691

Figure Lengend Snippet: Hepatic immune infiltration induced by conA. Hematoxylin and Eosin-stained liver sections obtained from conA-treated wild-type (A) and TβRI CA /Fsp1-Cre mice (B). Myeloperoxidase immunohistochemical staining of liver sections from conA-treated wild-type (C) and TβRI CA /Fsp1-Cre mice (D). CD3 immunohistochemical staining of liver sections from conA-treated wild-type (E) and TβRI CA /Fsp1-Cre mice (F). Quantification of myeloperoxidase-(G), and CD3-(F) positive cells in livers from PBS-treated and conA-treated mice (n = 6). Flow cytometry analysis of CD4 + cells (I), CD4 + IFN-γ + cells (J) and CD4 + IL-4 + cells (K) in livers from conA-treated wild-type and TβRI CA /Fsp1-Cre mice (n = 6). (L) Representative results of TCR-β and NK1.1 positivity by flow cytometry. (M) Quantitative analysis of TCR-β- and NK1.1-positive NKT cells by flow cytometry (n = 6). Data are shown as means ± SD. ∗ P < 0.05, ∗∗∗∗ P < 0.0001 between the two indicated groups.

Article Snippet: TβRI CA mice were kindly provided by Professor Laurent Bartholin, INSERM, France.

Techniques: Staining, Immunohistochemical staining, Flow Cytometry

Journal: Heliyon

Article Title: Macrophage expression of constitutively active TβRI alleviates hepatic injury in a mouse model of concanavalin A-induced autoimmune hepatitis

doi: 10.1016/j.heliyon.2025.e42691

Figure Lengend Snippet: M2 macrophage polarization was enhanced in livers of conA-treated TβRI CA /Fsp1-Cre mice. (A) Representative results of F4/80 and CD163 positivity in liver macrophages by flow cytometry. (B) Quantitative analysis of F4/80- and CD163-positive liver macrophages by flow cytometry (n = 6). Expression of CD206 (C) and CCR2 (D) in livers of PBS- and conA-treated wild-type and TβRI CA /Fsp1-Cre mice (n = 6). Data are shown as means ± SD. ∗ P < 0.05, ∗∗ P < 0.01 between the two indicated groups.

Article Snippet: TβRI CA mice were kindly provided by Professor Laurent Bartholin, INSERM, France.

Techniques: Flow Cytometry, Expressing

Journal: Heliyon

Article Title: Macrophage expression of constitutively active TβRI alleviates hepatic injury in a mouse model of concanavalin A-induced autoimmune hepatitis

doi: 10.1016/j.heliyon.2025.e42691

Figure Lengend Snippet: Gene expression analysis of liver macrophages from conA-treated TβRI CA /Fsp1-Cre mice. (A) Detection of Arg 1, Ym1 , and CD206 transcripts by qPCR performed on total RNA from liver macrophages of conA-treated mice. (B) Expression of H2-Aa and H2-k1 in liver macrophages from conA-treated wild-type and TβRI CA /Fsp1-Cre mice. (C) Expression of FOXO1 and IRF1 in liver macrophages from conA-treated wild-type and TβRI CA /Fsp1-Cre mice. (D) Quantification of CD1d mRNA expression levels in liver macrophages from conA-treated wild-type and TβRI CA /Fsp1-Cre mice. Data are shown as means ± SD of n = 5 per experimental group. ∗ P < 0.05, ∗∗ P < 0.001, ∗∗∗ P < 0.001 between the two indicated groups.

Article Snippet: TβRI CA mice were kindly provided by Professor Laurent Bartholin, INSERM, France.

Techniques: Gene Expression, Expressing

Journal: Scientific Reports

Article Title: Differential kinase activity of ACVR1 G328V and R206H mutations with implications to possible TβRI cross-talk in diffuse intrinsic pontine glioma

doi: 10.1038/s41598-020-63061-0

Figure Lengend Snippet: Dose response of SF8628 DIPG cell viability. ( a ) Single agent study of TβRI inhibitors. * p < 0.05, ** p < 0.01, *** p < 0.001, n = 4, unpaired t -test of highest dose vs. control. ( b ) panobinostat and (c) GSK-J4 tested in combination with 10 µM TβRI inhibitors. The relative cell viability of drug(s) treated samples over the DMSO control is shown. When, as in ( c ) for EW7197, the IC50 value is inconclusive from curve fitting, an apparent upper limit is estimated.

Article Snippet: The TβRI inhibitors EW7197 (vactosertib), LY3200882 and LY2157299 (galunisertib), SB525334, histone deacetylase inhibitor panobinostat and histone demethylase inhibitor GSK-J4 (all from Fisher Scientific, USA and manufactured by Cayman Chemicals) were administered as drug treatments to assess their effects on SF8628 cell growth.

Techniques: Control

Journal: Oncogene

Article Title: The TβRI promotes migration and metastasis through thrombospondin 1 and ITGAV in prostate cancer cells

doi: 10.1038/s41388-024-03165-3

Figure Lengend Snippet: a Exon 2 of TβRI was designed to be targeted by the mutant Cas9 enzyme. The mutant clone A9 contained a deletion of TβRI as detected by sequencing and the stop sequence formed by a frameshift mutation. b The mutant cell clones (A–E) and wild-type (WT) cells were examined by Cel-1 assay (Surveyor nuclease assay). c Mutant clones B5 and A9 and WT cells detected by RT-PCR. d , e Western blot and f qPCR were used to verify the TβRI-deficient cell lines. Results are shown as mean ± SEM, * p < 0.05, ** p < 0.01, Student’s t -test. g WT, A9, and HA - TβR1 reconstituted A9 cells were lysed and subjected to immunoblotting with antibodies TβRI, pSmad2, Smad2, and actin. h CAGA-luciferase reporter assay showing the TGFβ response of the indicated cell lines. Results are shown as mean ± SEM, * p < 0.05, *** p < 0.001, Student’s t -test.

Article Snippet: ML is a founder, shareholder, and board member of the company MetaCurUm Biotech AB that develops TβRI-based cancer therapies and biomarkers.

Techniques: Mutagenesis, Sequencing, Clone Assay, Nuclease Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Luciferase, Reporter Assay

Journal: Oncogene

Article Title: The TβRI promotes migration and metastasis through thrombospondin 1 and ITGAV in prostate cancer cells

doi: 10.1038/s41388-024-03165-3

Figure Lengend Snippet: a Gene-modified cell lines wild-type (WT), A9 (TβRI deficient), A9 HA-TβRI (A9 cells reconstituted with HA-TβRI), and A9 vector (empty vector as control) were subjected to immunoblotting with antibodies pSmad2, Smad2, TβRI, and actin. b CAGA-luciferase reporter assay showing the TGFβ response in the indicated cell lines. Results are shown as mean ± SEM, *** p < 0.001, Student’s t -test. c Expression of Smad7 and SERPINE1 in the indicated cell lines detected by qPCR. Results are shown as mean ± SEM, *** p < 0.001, Student’s t -test. d Microarray data showing the differentially expressed genes (DEGs) in the indicated cell lines with or without TGFβ treatment. e DEGs were annotated to the biological functions by DAVID Bioinformatics Resources. * p < 0.05, ** p < 0.01.

Article Snippet: ML is a founder, shareholder, and board member of the company MetaCurUm Biotech AB that develops TβRI-based cancer therapies and biomarkers.

Techniques: Modification, Plasmid Preparation, Control, Western Blot, Luciferase, Reporter Assay, Expressing, Microarray

Journal: Oncogene

Article Title: The TβRI promotes migration and metastasis through thrombospondin 1 and ITGAV in prostate cancer cells

doi: 10.1038/s41388-024-03165-3

Figure Lengend Snippet: a – c Expression of THBS1 and ITGAV detected by western blotting in the PC3U, DU-145, and A549 cell lines with or without a knockout or knockdown of TβRI. d Immunofluorescent (IF) staining of THBS1 and pSmad2 in WT and A9 cells in a wound-healing assay with or without TGFβ treatment. Scale bar, 20 μm. e Invasion assay showing the invasive capacity of PC3U cells with or without knockout of TβRI or with or without TGFβ treatment for 48 h. Scale bar, 50 μm. f Western blotting showing the overexpression of THBS1 in A9 cells. g Invasion assay showing the invasive capacity of WT, A9, and A9 cells overexpressed THBS1. Scale bar, 50 μm. h The invading cells were quantified by OD value. Shown as mean ± SD, * p < 0.05. i IF staining of THBS1 and Paxillin in PC3U cells treated with TGFβ for 6 h. Scale bar, 20 μm. j IF staining of THBS1, Paxillin, and Phalloidin in PC3U cells treated with TGFβ for 6 h. Scale bar, 20 μm.

Article Snippet: ML is a founder, shareholder, and board member of the company MetaCurUm Biotech AB that develops TβRI-based cancer therapies and biomarkers.

Techniques: Expressing, Western Blot, Knock-Out, Knockdown, Staining, Wound Healing Assay, Invasion Assay, Over Expression

Journal: Oncogene

Article Title: The TβRI promotes migration and metastasis through thrombospondin 1 and ITGAV in prostate cancer cells

doi: 10.1038/s41388-024-03165-3

Figure Lengend Snippet: a Co-IF staining of HA, THBS1, and ITGAV in the wild-type (WT), A9 (TβRI deficient), A9 HA-TβRI (A9 cells reconstituted with HA-TβRI) cell lines with or without TGFβ treatment. Scale bar, 20 μm. b Co-IP was performed to detect the interaction of HA-TβRI with THBS1 and ITGAV. c The correlation of TβRI and THBS1 expression, TβRI and ITGAV expression in prostate cancer (TCGA database). d The interaction of THBS1 and ITGAV was detected in a wound-healing assay by in situ PLA. Scale bar, 50 μm. e Invasion assay showing that knockdown of THSB1 prevented the invasion capacity of PC3U cells treated with TGFβ. Scale bar, 50 μm. f The invading cells were quantified by OD value. Shown as mean ± SD, * p < 0.05. g Western blot showing the expression of THSB1 and ITGAV in PC3U cells with THSB1 knockdown.

Article Snippet: ML is a founder, shareholder, and board member of the company MetaCurUm Biotech AB that develops TβRI-based cancer therapies and biomarkers.

Techniques: Staining, Co-Immunoprecipitation Assay, Expressing, Wound Healing Assay, In Situ, Invasion Assay, Knockdown, Western Blot

Journal: Oncogene

Article Title: The TβRI promotes migration and metastasis through thrombospondin 1 and ITGAV in prostate cancer cells

doi: 10.1038/s41388-024-03165-3

Figure Lengend Snippet: a Wild-type (WT) PC3U and TβRI-deficient A9 cells were injected into the prostates of mice. Tumors and sentinel lymph nodes were obtained from the orthotopic xenograft mouse model after 4 weeks. b The tumor weight was quantified in both the WT ( n = 9) and A9 ( n = 9) groups. Results are shown as mean ± SEM, *** p < 0.001, one-way ANOVA. c The metastatic lymph nodes were quantified in both the WT ( n = 9) and A9 ( n = 6) groups. Results are shown as mean ± SEM, *** p < 0.001, one-way ANOVA. d Representative images of THBS1 staining in the WT or A9 tumors and lymph metastasis. Scale bar, 500 μm. Left: the black arrow shows the lymph vessels in the tumor-invasive front, and the red arrow shows tumor cells infiltrating into the adjacent muscles. Right: the black arrows show the remaining lymphatic cells in the sentinel lymph nodes. Scale bar, 100 μm. e Representative images of THBS1 staining in the normal adjacent prostate tissue, lower malignant prostatic adenocarcinoma (GS ≤ 6), and higher malignant prostatic adenocarcinoma (GS > 6). f THBS1 expression was examined on tissue microarray sections with 192 samples. Immunoreactivity (IR) was scored as negative (IR = 0), weak (IR = 1), moderate (IR = 2), or strong (IR = 3) **** p < 0.0001, Tukey’s multiple comparisons test. g A proteomic database was used to analyze the THBS1 protein level in the control adjacent prostate tissue ( n = 8), localized tumor ( n = 28), and bone metastasis ( n = 22). **** p < 0.0001, one-way ANOVA. h Representative images for the IF staining of THBS1 and panCK AE1/3 in the human primary and bone metastatic tumors. Scale bar, 50 μm.

Article Snippet: ML is a founder, shareholder, and board member of the company MetaCurUm Biotech AB that develops TβRI-based cancer therapies and biomarkers.

Techniques: Injection, Staining, Muscles, Expressing, Microarray, Control