Journal: PLoS ONE

Article Title: CXCL12/CXCR4 Axis Activation Mediates Prostate Myofibroblast Phenoconversion through Non-Canonical EGFR/MEK/ERK Signaling

doi: 10.1371/journal.pone.0159490

Figure Lengend Snippet: ( A ) N1 fibroblasts were treated with vehicle (DMSO) or EGFR inhibitor (AG1478, 500 nM) prior to CXCL12 treatment. Phosphorylation of EGFR, Akt, Smad and ERK1/2 were assessed via western blot. ( B ) qRT-PCR analysis of myofibroblast marker expression after CXCL12 (100pM) treatment in the presence or absence of AG1478 (500 nM). Expression levels of α-smooth muscle actin (ACTA2) and collagen 1α1 (COL1α1) were analyzed over the course of 24 hours of treatment. Treatment with AG1478 reduced or ablated the CXCL12/CXCR4 -mediated stimulation of both the ACTA2 and COL1α1 genes. ( C ) Western blot analysis of fibroblast markers, α-smooth muscle actin and collagen 1, at 24 and 48 hours after CXCL12 treatment. Secretion and incorporation of collagen 1 was inhibited in the presence of AG1478, α-smooth muscle actin production was inhibited as well. ( D ) Signal intensity quantification for 7 and collagen 1 western blots. * = p-value < 0.05. Error bars, SE.

Article Snippet: Primary antibodies were diluted in blocking solution and included 1:200 dilution FITC-conjugated mouse monoclonal anti-α-smooth muscle actin (α SMA), and 1:100 dilution biotin-conjugated rabbit polyclonal anti-collagen type 1 (COL1) (Rockland Immunochemicals, Gilbertsville, PA).

Techniques: Phospho-proteomics, Western Blot, Quantitative RT-PCR, Marker, Expressing

Journal: PLoS ONE

Article Title: CXCL12/CXCR4 Axis Activation Mediates Prostate Myofibroblast Phenoconversion through Non-Canonical EGFR/MEK/ERK Signaling

doi: 10.1371/journal.pone.0159490

Figure Lengend Snippet: ( A ) Immunofluorescence analysis of N1 fibroblasts left untreated (-CXCL12) or treated with 100pM CXCL12 (CXCL12) for 48 hours in the absence or presence of CXCR4 (250 μM AMD3100), EGFR (250 uM AG1478), ALK-5 (TGFβRII) (20μM A-83-01) small molecular inhibitors or an antibody against TGFβRII (200 ng/ml TGFβ MAb). Figure depicts photomicrographs images of cells stained for α-smooth muscle actin (green) or collagen 1 (red) proteins or DAPI (blue nuclear stain); orange color indicates α-smooth muscle actin and collagen 1 colocalization in the merged image. ( B ) Western blot analysis of fibroblast markers, α-smooth muscle actin and collagen 1 at 24, 48 and 72 hours after CXCL12 (100pM) of scramble, anti-CXCR4, and anti-TGFβRI siRNAs transfected fibroblasts. CXCL12-driven expression of myofibroblasts markers does not require the presence or activation of TGFβRI. ( C ) Quantification of western blot images for myofibroblast markers.

Article Snippet: Primary antibodies were diluted in blocking solution and included 1:200 dilution FITC-conjugated mouse monoclonal anti-α-smooth muscle actin (α SMA), and 1:100 dilution biotin-conjugated rabbit polyclonal anti-collagen type 1 (COL1) (Rockland Immunochemicals, Gilbertsville, PA).

Techniques: Immunofluorescence, Staining, Western Blot, Transfection, Expressing, Activation Assay

Journal: Bioactive Materials

Article Title: 3D bioprinted multi-layered cell constructs with gradient core-shell interface for tendon-to-bone tissue regeneration

doi: 10.1016/j.bioactmat.2024.10.002

Figure Lengend Snippet: Preparation of bioink-T for the fabrication of tendon tissue. (A) Rheological properties (G′) for the tendon-derived dECM methacrylate (TdECMA)-based bioink containing diverse concentrations of the PVA (50, 100, and 150 mg/mL) (n = 6). (B) Live/dead (at 3 and 7 d) and DAPI/F-actin (green) (at 7 d) images and quantified (C) cell viability, (D) orientation factor, and F-actin positive area for bioprinted structures using the TdECMA/PVA bioinks (n = 6). (E) Scanning electron microscope (SEM), collagen type-I (COL1), and DAPI/F-actin (at 7 d) images for the tendon structures fabricated with TdECMA naive bioink (TS-1) and TdECMA/PVA-based bioinks with (TS-3) and without (TS-2) transforming growth factor beta (TGF-β). (F) Quantified orientation factor of the actin filament (n = 6). (G) DAPI/TNMD images and quantitatively analyzed (H) TNMD positive area and (I) tenogenesis-related gene expression for the bioprinted tendon constructs (n = 6). The synergistic effects of cellular alignment and tenogenic molecules in TS-3 significantly improved the differentiation of hASCs into the tenogenic lineage compared to TS-1 and TS-2. All values are presented as the mean ± SD (∗∗ p < 0.01 and ∗∗∗ p < 0.001).

Article Snippet: Then, the prepared cells were incubated with a rabbit anti-collagen type I (COL1) (5 μg/mL in DPBS; Invitrogen), a mouse anti-osteopontin (OPN) (5 μg/mL in DPBS; Invitrogen), a rabbit anti-tenomodulin (TNMD) (5 μg/mL in DPBS; Invitrogen), and a rat anti-aggrecan (ACAN) (5 μg/mL in DPBS; Invitrogen) primary antibodies overnight at 4 °C, followed by staining the samples using Alexa Fluor 488- mouse, 594-conjugated rabbit, or 647-conjugated rat secondary antibodies (1:50 in DPBS; Invitrogen) for 1 h at 37 °C.

Techniques: Derivative Assay, Microscopy, Gene Expression, Construct

Journal: Bioactive Materials

Article Title: 3D bioprinted multi-layered cell constructs with gradient core-shell interface for tendon-to-bone tissue regeneration

doi: 10.1016/j.bioactmat.2024.10.002

Figure Lengend Snippet: Immunohistochemical imaging of the bioprinted TBI construct in rabbit rotator cuff tear model. (A) COL1, COL3, platelet and endothelial cell adhesion molecule 1 (PECAM-1), and vascular endothelial growth factor (VEGF) immunohistochemical staining images at 8 weeks post-implantation. Semi-quantitatively estimated score of (B) COL1, (C) COL3, (D) PECAM-1, and (E) VEGF (n = 4). (F) Immunohistochemical staining images and (G) semi-quantitatively analyzed score of HLA Class 1 ABC (HLA-ABC) (n = 4). Red and blue arrows indicate the HLA-ABC-positive and -negative cells, respectively. The most efficient regenerative capacities, including tendon-to-bone tissue integration and vascular formation, were observed in the rabbit RCT model with the transplantation of the Exp bioconstructs, validated by immunohistochemical imaging analyses. All values are presented as the mean ± SD (∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001).

Article Snippet: Then, the prepared cells were incubated with a rabbit anti-collagen type I (COL1) (5 μg/mL in DPBS; Invitrogen), a mouse anti-osteopontin (OPN) (5 μg/mL in DPBS; Invitrogen), a rabbit anti-tenomodulin (TNMD) (5 μg/mL in DPBS; Invitrogen), and a rat anti-aggrecan (ACAN) (5 μg/mL in DPBS; Invitrogen) primary antibodies overnight at 4 °C, followed by staining the samples using Alexa Fluor 488- mouse, 594-conjugated rabbit, or 647-conjugated rat secondary antibodies (1:50 in DPBS; Invitrogen) for 1 h at 37 °C.

Techniques: Immunohistochemical staining, Imaging, Construct, Staining, Transplantation Assay

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: MTFR2-Mediated Fission Drives Fatty Acid and Mitochondrial Co-Transfer from Hepatic Stellate Cells to Tumor Cells Fueling Oncogenesis.

doi: 10.1002/advs.202416419

Figure Lengend Snippet: Figure 1. MTFR2-mediated mitochondrial fission in aHSCs enhances HCC progression. A–D) Histological analysis of para-tumor and tumor margin regions from HCC patient and mouse orthotopic HCC model. A) Hematoxylin and eosin (H&E) staining and immunofluorescence (IF) for COL1 (green), 𝛼-SMA (red), and GFAP (purple, indicating HSC) in the margin area and para-tumor tissue from an HCC patient. B) Schematic illustration indicating tissue collection sites for (A)—the margin area and para-tumor tissue. C) Schematic for the tissue collection shown in (D), representing areas in a mouse orthotopic HCC model. D) H&E and IF staining of liver tissues to compare tumor margin and para-tumor regions. Scale bars: 100 μm. E) Immunoelectron

Article Snippet: Antibodies used for western blot, immunohistochemistry (IHC), and immunofluorescence: anti-collagen type I (COL1) polyclonal antibody (Catalog No. 14695-1- AP) was purchased from Proteintech Group, Inc. (Chicago, USA) and anti-alpha smooth muscle actin (α-SMA) mouse monoclonal antibody (Catalog No. BM0002) was purchased from Boster Biological Co. Ltd. (Wuhan, China).

Techniques: Staining

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: MTFR2-Mediated Fission Drives Fatty Acid and Mitochondrial Co-Transfer from Hepatic Stellate Cells to Tumor Cells Fueling Oncogenesis.

doi: 10.1002/advs.202416419

Figure Lengend Snippet: Figure 6. MTFR2 in HSCs drives tumor progression in vivo. A) Schematic illustration showing the experimental setup for tumor induction with MHCC- 97H cells and different HSC conditions. B) Graph depicting tumor volume over time for MHCC-97H, MHCC-97H + WT HSC, and MHCC-97H + KD HSC groups (Data are represented as mean ± SD, n = 6, two-way ANOVA was performed). C) Images of tumor nodules formed from MHCC-97H cells co- cultured with either wild-type (WT) HSCs or knockdown (KD) HSCs. D) Analysis of the final tumor body weight ratio (%) (Data are represented as mean ± SD, n = 6, one-way ANOVA was performed). E) Histological assessment via hematoxylin and eosin (H&E) staining, and immunohistochemical analysis of Ki67, 𝛼-SMA, and COL1 in tumor sections. Scale bars: 100 μm. F) Immunofluorescence staining of various proteins (GFAP, MTFR2, DRP1, ACC1) in tumor sections, indicating expression patterns. Scale bars: 100 μm. G) Western blot analysis of CPT1A and CPT2 expression. H) IF staining of CPT1A in tumor tissues of different groups. GFP-HSC was shown to distinguish HSCs. Scale bars: 50 μm. I) Fluorescent images showing the mito-mcherry labeled mitochondria from HSCs in GPC3 positive MHCC-97H cells (white arrows). Scale bars: 30 μm.

Article Snippet: Antibodies used for western blot, immunohistochemistry (IHC), and immunofluorescence: anti-collagen type I (COL1) polyclonal antibody (Catalog No. 14695-1- AP) was purchased from Proteintech Group, Inc. (Chicago, USA) and anti-alpha smooth muscle actin (α-SMA) mouse monoclonal antibody (Catalog No. BM0002) was purchased from Boster Biological Co. Ltd. (Wuhan, China).

Techniques: In Vivo, Cell Culture, Knockdown, Staining, Immunohistochemical staining, Expressing, Western Blot, Labeling

Journal: Stem Cells International

Article Title: Retinoic Acid Signal Negatively Regulates Osteo/Odontogenic Differentiation of Dental Pulp Stem Cells

doi: 10.1155/2020/5891783

Figure Lengend Snippet: Blocking RA signal enhances the expression of osteo/odontogenic related factors from human DPSCs in vivo. Immunohistochemical (IHC) staining reveals expression levels of DSPP (a, b), OCN (d, e), and COL-1 (g, h). Qualitative measurements of IHC staining show expressions of DSPP (c), OCN (f), and COL-1 (i) are enhanced significantly by BMS 493 treatment. All error bars mean the SD ( n = 5). ∗∗∗ P < 0.001 and ∗∗∗∗ P < 0.0001.

Article Snippet: The primary antibodies used here included those against dentin sialophosphoprotein (DSPP) (Cat. No. ab216892, Abcam, Cambridge, UK), osteocalcin (OCN) (Cat No. ab13418, Abcam, Cambridge, UK), and collagen type I (COL-1) (Cat No. NB600–408, Novus Biological Centennial, USA).

Techniques: Blocking Assay, Expressing, In Vivo, Immunohistochemical staining, Immunohistochemistry

Journal: Stem Cells International

Article Title: Retinoic Acid Signal Negatively Regulates Osteo/Odontogenic Differentiation of Dental Pulp Stem Cells

doi: 10.1155/2020/5891783

Figure Lengend Snippet: The schematic diagram of the effect of RA on DPSCs. Activation of RA inhibits osteo/odontogenic differentiation of DSPCs and the expression levels of osteo/odontogenic related factors ALP, COL-1, OPN, OCN, and DSPP by downregulating the expression of OSX.

Article Snippet: The primary antibodies used here included those against dentin sialophosphoprotein (DSPP) (Cat. No. ab216892, Abcam, Cambridge, UK), osteocalcin (OCN) (Cat No. ab13418, Abcam, Cambridge, UK), and collagen type I (COL-1) (Cat No. NB600–408, Novus Biological Centennial, USA).

Techniques: Activation Assay, Expressing