Journal: EBioMedicine

Article Title: Autocrine CTHRC1 activates hepatic stellate cells and promotes liver fibrosis by activating TGF-β signaling

doi: 10.1016/j.ebiom.2019.01.009

Figure Lengend Snippet: CTHRC1 activates both TGF-β and Wnt signaling, while the promotive effect of CTHRC1 on HSC activation is mainly dependent on TGF-β signaling. A. The expression of α-SMA in primary HSCs after 7 days isolated from WT and CTHRC1 −/− mice. B. The expression of α-SMA in primary rat HSCs after 4 days, which were treated with vehicle, 20 nM rCTHRC1 protein alone, and 20 nM rCTHRC1 protein plus CTHRC1 mAb or IgG. C. Western blotting analysis of phosphorylation of Smad2, Smad3, JNK, total Smad4 and expression of Wnt5a in five liver tissues of WT or CTHRC1 −/− mice intraperitoneally injected with CCl 4 . GAPDH was the loading control. The densitometry of p-Smad2/Smad2 was shown below. D. Phosphorylation of Smad2, Smad3, JNK, total Smad4 and expression of Wnt5a in primary rat HSCs, which were treated with vehicle, 20 nM rCTHRC1 protein, and 20 nM rCTHRC1 protein plus CTHRC1 mAb or IgG for 1, 3, 5 days, individually. GAPDH was the loading control. The densitometry of p-Smad2/Smad2 was shown below. E and F. Representative immunofluorescence images of α-SMA (green in E, red in F) in primary rat HSCs after 4 days, which were treated with vehicle, 20 nM rCTHRC1 protein, and 20 nM rCTHRC1 protein plus neutralizing antibodies or inhibitor as follows: TGFBR2 neutralizing antibody or TGF-β receptor inhibitor (E), Wnt5a or Wnt3a neutralizing antibody (F). Nuclei are stained with DAPI (blue). Scale bars, 50 μm. G. The expression of α-SMA in primary rat HSCs after 4 days, which were treated with vehicle, 20 nM rCTHRC1 protein, and 20 nM rCTHRC1 protein plus TGFBR2 neutralizing antibody or TGF-β receptor inhibitor. GAPDH was the loading control. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Recombinant CTHRC1 and/or TGFBR2 neutralizing antibody (R&D, AF-241-NA), TGF-βR inhibitor, LY2109761 (SELLECK, S2704) were added in the medium simultaneously.

Techniques: Activation Assay, Expressing, Isolation, Western Blot, Injection, Immunofluorescence, Staining

Journal: Stem Cells and Development

Article Title: Resistin-Like Molecule ? Stimulates Proliferation of Mesenchymal Stem Cells While Maintaining Their Multipotency

doi: 10.1089/scd.2012.0192

Figure Lengend Snippet: Effect of RELMα on osteogenic and myogenic differentiation of MSCs. (A) Osteogenic differentiation was stimulated for 3 weeks with osteogenic medium, cells were stained with Alizarin Red for calcium phosphate deposits. The staining was quantified using ImageJ softwate (*P=0.2). (B) Western blot shows the expression of osteopontin. (C) Myogenic differentiation was induced with 10 ng/mL transforming growth factor β1 in growth medium for 5 days. Western blot shows the expression of smooth muscle actin and fibronectin as markers of myofibroblasts.

Article Snippet: The following antibodies were used: phospho-p38 MAPK (Thr180/Tyr182) mouse mAb, phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) rabbit mAb, Egr-1 rabbit mAb, phospho-Akt, phospho-p65 (Ser536) rabbit mAb, adiponectin rabbit mAb, and β-Tubulin rabbit mAb (Cell Signaling Technology); β-actin mouse mAb clone AC-15 and GAPDH mouse mAb (Sigma-Aldrich); RELMα goat antibody (R & D Systems); osteopontin mouse mAb (Abcam), fibronectin mouse mAb, fluorescein isothiocyanate (FITC)-labeled rat anti-mouse Ly-6A/E (Sca-1) Ab, and FITC-labeled rat anti-mouse CD45 Ab (BD Biosciences).

Techniques: Staining, Western Blot, Expressing