Journal: Bioengineering

Article Title: All Trans-Retinoic Acids Facilitate the Remodeling of 2D and 3D Cultured Human Conjunctival Fibroblasts

doi: 10.3390/bioengineering9090463

Figure Lengend Snippet: Effects of ATRA toward the gene expression of ECMs of the TGF-β2 untreated or treated HconF cells. Two-dimensional and three-dimensional HconF cells untreated and treated with TGF-β2 (TGFβ, 5 ng/mL) were subjected to qPCR analysis in the presence or absence of 1 μM ATRA and the mRNA expressions in ECMs, including COL1 , COL4 , COL6 , FN, and aSMA, were evaluated. All experiments were performed in triplicate using 3 different confluent 6-well dishes (2D) or 15 freshly prepared 3D HconF spheroids (3D) in each experimental condition. Data are expressed as the mean ± standard error of the mean (SEM). * p < 0.05, ** p < 0.01 and *** p < 0.005; ANOVA followed by a Tukey’s multiple comparison test.

Article Snippet: They were then sequentially treated with (1) 1:200 dilutions of primary antibodies; an anti-human COL1 (#600-401-103-0.1, ROCKLAND Antibodies & Assays, Limerick, PA, USA), COL4 (#600-401-106S, ROCKLAND Antibodies & Assays, Limerick, PA, USA), COL6 (#009-001-108, ROCKLAND Antibodies & Assays, Limerick, PA, USA), or FN (#A-11, Santa Cruz Biotechnology, Inc., Dallas, TX, USA) rabbit antibody at 4 °C overnight, (2) washed 3 times with PBS for 1 h each, (3) 1:1000 dilutions of a secondary antibody; a goat anti-rabbit IgG (488 nm, #A27034, Invitrogen, Waltham, MA, USA) with phalloidin (594 nm, 1:1000 dilutions, # 17466-45-4, Cayman Chemical, Ann Arbor, MI, USA) and DAPI (1:1000 dilutions, # D523-10, DOJINDO, Osaka, Japan) for 3 hrs, and (4) mounting ProLong Gold Antifade Mountant with a cover glass.

Techniques: Gene Expression, Comparison

Journal: Cell Reports Physical Science

Article Title: Multiscale hydrogel regulates mesenchymal stem cell fate for bone regeneration

doi: 10.1016/j.xcrp.2024.102181

Figure Lengend Snippet: Figure 8. Immunolabeling of ADSC-loaded locally stiffness-enhanced macroporous hydrogel during calvarial healing IHC detection of osteogenic differentiation markers collagen type I (Col1, A), and OCN (B) at post-operative weeks 4 and 8. (C and D) Quantitative analysis of immunohistochemistry (n = 3; *p < 0.05; **p < 0.01; ***p < 0.001).

Article Snippet: After sectioning, the cells were treated with specific antibodies as follows: rabbit anti-ALP (1:100; bsm-52252R, Bioss Antibodies, Woburn, MA), OCN polyclonal antibody (1:200; 23418-1-AP, Proteintech, Rosemont, IL), anti-Col1 antibody (1:200; BA0325, Boster Bio, Pleasanton, CA), andmonoclonal anti-YAP1 antibody produced in mice (1:200; SAB1404823, Sigma-Aldrich); these were incubated at 4 C for 12–16 h. After incubation of the fluorochrome-conjugated secondary antibodies under protected light conditions, the nuclei and cytoskeleton were stained with DAPI (28718-90-3, Solarbio) and actin (1:100; bs-1571R, Bioss Bio).

Techniques: Immunolabeling, Immunohistochemistry

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: International Journal of Molecular Sciences

Article Title: Interleukin-17A Orchestrates Lung Injury and Remodeling Through p53 and uPA System Crosstalk

doi: 10.3390/ijms27041841

Figure Lengend Snippet: Regulation of BLM-LI and TSE-LI by IL-17A. ( A ) Lung sections of WT mice exposed to saline or BLM for 24–72 h were subjected to IHC staining for IL-17A antigen ( i ). AECs isolated from WT and IL-17A-deficient mice exposed to saline or BLM were analyzed for apoptosis and Sirt1 expression by Western blotting (WB) three days later ( ii ). Lung sections of WT and IL-17A −/− mice exposed to saline or BLM were subjected to Masson’s trichrome staining 21 days later ( iii ). Whole lung homogenates of WT and IL-17A −/− mice exposed to saline or BLM as in were analyzed for total HYP content ( iv ) or for Col-1 and FN by WB 21 days after BLM ( v ). ( B ) WT, p53 −/− and PAI-1 −/− mice were kept in ambient air or exposed to passive TS for 20 weeks. Total lung homogenates of control mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice were analyzed for IL-17A and IL-17RA protein by WB ( i ). Total RNA from mice kept in ambient air, TSE WT mice, and p53 −/− or PAI-1 −/− mice was analyzed for IL-17A mRNA ( ii ). AECs from TSE WT and IL-17A −/− mice were analyzed for p53, ACp53, cleaved (Cl.) caspase-3/caspase-3 and Sirt1 by WB ( iii ). AECs from lungs of IL-17A −/− mice kept in ambient air or TSE, as well as mice treated with or without CSP7 or CP, were analyzed for telomere length by qPCR ( iv ). AECs from lungs of IL-17A −/− mice treated as in ( B(iv )) were immunoblotted for the listed proteins ( v ). Lung sections of IL-17A −/− mice treated as in ( B ( iv )) were subjected to IHC for telomere binding shelterin complex (TRF1 and TRF2) proteins ( vi ).

Article Snippet: 14 , Anti-Col-1 mAb , Southern Biotech , #1340-01 , 1:1000 , .

Techniques: Saline, Immunohistochemistry, Isolation, Expressing, Western Blot, Staining, Control, Binding Assay