Journal: Frontiers in Pharmacology

Article Title: Latent Transforming Growth Factor-β Binding Protein-2 Regulates Lung Fibroblast-to-Myofibroblast Differentiation in Pulmonary Fibrosis via NF-κB Signaling

doi: 10.3389/fphar.2021.788714

Figure Lengend Snippet: LTBP2 silencing inhibits TGFβ1-induced differentiation of fibroblasts in vitro . (A) Quantitative RT-PCR and western blot analysis of α-SMA, fibronectin, and collagen I in HFL1 cells transfected with scrambled shRNA or LTBP2 shRNA followed by PBS or TGF-β1 stimulation. GAPDH was detected as the internal control. ** p < 0.01 vs. scr shRNA, # p < 0.05, ## p < 0.01 vs. scr shRNA + TGFβ1 (B) Representative immunofluorescence staining of α-SMA (red) in HFL1 cells. Nuclei are stained with DAPI (blue). Scale bar = 20 μm. (C) Representative immunofluorescence staining of collagen I (red) in HFL1 cells. Nuclei are stained with DAPI (blue). Scale bar = 20 μm. Data are represented as means ± SD, n = 3. Statistical analysis was performed by one-way ANOVA.

Article Snippet: HFL1 cells were pretreated with BAY 11-7082 (MedChem Express, NJ, United States) for 1 h followed by transfection with LTBP2 plasmids.

Techniques: In Vitro, Quantitative RT-PCR, Western Blot, Transfection, shRNA, Control, Immunofluorescence, Staining

Journal: Frontiers in Pharmacology

Article Title: Latent Transforming Growth Factor-β Binding Protein-2 Regulates Lung Fibroblast-to-Myofibroblast Differentiation in Pulmonary Fibrosis via NF-κB Signaling

doi: 10.3389/fphar.2021.788714

Figure Lengend Snippet: LTBP2 silencing suppresses the activation of NF-κB signaling induced by TGFβ1. (A) Representative immunofluorescence staining of phospho-NF-κB p65 (ser536) (red) in HFL1 cells transfected with scrambled shRNA or LTBP2 shRNA followed by PBS or TGF-β1 stimulation. Nuclei are stained with DAPI (blue). Scale bar = 20 μm. (B) Western blot analysis of nuclear NF-κB p65 and nuclear phospho-NF-κB p65 (ser536) in HFL1 cells. Lamin B was detected as the internal control. ** p < 0.01 vs. scr shRNA, # p < 0.05, ## p < 0.01 vs. scr shRNA + TGFβ1. Data are represented as means ± SD, n = 3. Statistical analysis was performed by one-way ANOVA.

Article Snippet: HFL1 cells were pretreated with BAY 11-7082 (MedChem Express, NJ, United States) for 1 h followed by transfection with LTBP2 plasmids.

Techniques: Activation Assay, Immunofluorescence, Staining, Transfection, shRNA, Western Blot, Control

Journal: Frontiers in Pharmacology

Article Title: Latent Transforming Growth Factor-β Binding Protein-2 Regulates Lung Fibroblast-to-Myofibroblast Differentiation in Pulmonary Fibrosis via NF-κB Signaling

doi: 10.3389/fphar.2021.788714

Figure Lengend Snippet: LTBP2 overexpression-induced lung fibroblast-to-myofibroblast differentiation depends on NF-κB phosphorylation and nuclear translocation in vitro . Western blot analysis of α-SMA, fibronectin, and collagen I in HFL1 cells transfected with LTBP2 cDNA and/or BAY 11-7082 or vector. GAPDH was detected as the internal control. ## p < 0.01 vs. vector. * p < 0.05, ** p < 0.01 vs. OE-LTBP2. Data are represented as means ± SD, n = 3. Statistical analysis was performed by one-way ANOVA.

Article Snippet: HFL1 cells were pretreated with BAY 11-7082 (MedChem Express, NJ, United States) for 1 h followed by transfection with LTBP2 plasmids.

Techniques: Over Expression, Phospho-proteomics, Translocation Assay, In Vitro, Western Blot, Transfection, Plasmid Preparation, Control

Journal: Stem Cell Research & Therapy

Article Title: Pre-incubation with hucMSC-exosomes prevents cisplatin-induced nephrotoxicity by activating autophagy

doi: 10.1186/s13287-016-0463-4

Figure Lengend Snippet: Human umbilical cord-derived mesenchymal stem cell exosomes ( hucMSC-Ex ) prevent deterioration of renal function in vivo. a Representative images of renal histology (200×, scale bar = 50 μm). b The histomorphological score. c Immunohistochemical analysis of PCNA expression in kidney tissues (200×, scale bar = 50 μm). d Percentage of proliferating cell nuclear antigen ( PCNA )-positive cells. e TUNEL assay for apoptotic cells in rat renal tissues (200×, scale bar = 50 μm). f Percentage of apoptotic cells. Serum creatinine ( Cr ) ( g ) and blood urea nitrogen ( BUN ) ( h ) levels were measured at different times after cisplatin injection. Data are expressed as mean ± SD. * P < 0.05, n = 6. Rats treated as mentioned for the in vivo rat model were divided into four groups as follows: 1) control; 2) phosphate-buffered saline ( PBS ); 3) hucMSC-Ex; and 4) human fetal lung fibroblast-1exosomes ( HFL1-Ex ). All animals were sacrificed at 3 days after cisplatin injection

Article Snippet: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining was used to detect the apoptotic cells. hucMSCs, HFL1 cells, NRK-52E cells, and the kidney tissues were examined using the TdT-FragEL DNA fragmentation detection kit according to the manufacturer's protocol (Boster, China).

Techniques: Derivative Assay, In Vivo, Immunohistochemical staining, Expressing, TUNEL Assay, Injection, Control, Saline

Journal: Stem Cell Research & Therapy

Article Title: Pre-incubation with hucMSC-exosomes prevents cisplatin-induced nephrotoxicity by activating autophagy

doi: 10.1186/s13287-016-0463-4

Figure Lengend Snippet: Human umbilical cord-derived mesenchymal stem cell exosomes ( hucMSC-Ex ) inhibit cisplatin-induced apoptosis and secretion of inflammatory cytokines in NRK-52E cells in vitro. a NRK-52E cells were stained with JC-1 dye and observed under fluorescent microscopy (JC-1 produces red fluorescence within the mitochondria as JC-1-aggregates while it emits green fluorescence when it leaks into the cytoplasm as JC-1-monomers; the shift between green and red is proportional to the mitochondrial membrane potential change) (200×, scale bar = 50 μm). b TUNEL assay was performed to detect the apoptotic cells (200×, scale bar = 50 μm). c Percentage of apoptotic cells. ** P < 0.01, n = 3. d The expression of proliferating cell nuclear antigen ( PCNA ), BCL-XL, BCL-2, Bax, Cytochrome-c, cleaved Caspase-3, and GAPDH proteins was evaluated by Western blot. e Immunohistochemical analysis of tumor necrosis factor alpha ( TNF-α ) expression (200×, scale bar = 50 μm). f The expression of interleukin 1-β ( IL1-β ) and GAPDH proteins was evaluated by Western blot. NRK-52E cells treated as mentioned for the in vitro experiment were divided into four groups: 1) control; 2) phosphate-buffered saline ( PBS ); 3) hucMSC-Ex; and 4) human fetal lung fibroblast-1exosomes ( HFL1-Ex )

Article Snippet: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining was used to detect the apoptotic cells. hucMSCs, HFL1 cells, NRK-52E cells, and the kidney tissues were examined using the TdT-FragEL DNA fragmentation detection kit according to the manufacturer's protocol (Boster, China).

Techniques: Derivative Assay, In Vitro, Staining, Microscopy, Fluorescence, Membrane, TUNEL Assay, Expressing, Western Blot, Immunohistochemical staining, Control, Saline

Journal: Stem Cell Research & Therapy

Article Title: Pre-incubation with hucMSC-exosomes prevents cisplatin-induced nephrotoxicity by activating autophagy

doi: 10.1186/s13287-016-0463-4

Figure Lengend Snippet: Human umbilical cord-derived mesenchymal stem cell exosomes ( hucMSC-Ex ) prevent apoptosis and secretion of inflammatory cytokines by activating autophagy in vivo. a Representative images of renal histology (200×, scale bar = 50 μm). b The histomorphological score. c Immunohistochemical analysis of PCNA expression in kidney tissues (200×, scale bar = 50 μm). d Percentage of proliferating cell nuclear antigen ( PCNA )-positive cells. e TUNEL assay for apoptotic cells in rat renal tissues (200×, scale bar = 50 μm). f Percentage of apoptotic cells. g The expression of LC3B, BCL-2, Bax, IL1-β, and GAPDH proteins in the cisplatin-induced rat model were measured by Western blot. The serum inflammatory cytokines ( h ) tumor necrosis factor alpha ( TNF-α ), ( i ) interleukin-1β ( IL1-β ), and ( j ) interleukin-6 ( IL6 ) were measured via Luminex assay. Data are expressed as mean ± SD. * P < 0.05, n = 6. Rats were divided into six groups as follows: 1) control; 2) phosphate-buffered saline ( PBS ); 3) hucMSC-Ex; 4) hucMSC-Ex + 3-methyladenine ( 3MA ); 5) 3MA; and 6) rapamycin ( Rapa )

Article Snippet: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining was used to detect the apoptotic cells. hucMSCs, HFL1 cells, NRK-52E cells, and the kidney tissues were examined using the TdT-FragEL DNA fragmentation detection kit according to the manufacturer's protocol (Boster, China).

Techniques: Derivative Assay, In Vivo, Immunohistochemical staining, Expressing, TUNEL Assay, Western Blot, Luminex, Control, Saline

Journal: Frontiers in Oncology

Article Title: Mining of RNA Methylation-Related Genes and Elucidation of Their Molecular Biology in Gallbladder Carcinoma

doi: 10.3389/fonc.2021.621806

Figure Lengend Snippet: Top 25 hub genes identified in PPI network for DEGs.

Article Snippet: Primary antibodies against the following proteins were purchased: FGA (1:50, Boster, China), F2 (1:50, Boster, China), CFH (1:50, Boster, China), PIPOX (1:50, Absin, China), ITIH4 (1:50, Proteintech, USA), GNMT (1:50, Proteintech, USA), MAT1A (1:50, Fine Biotech, China), MTHFD1 (1:50, Proteintech, USA), HPX (1:50, Boster, China), CTH (1:50, Boster, China), CFHR3 (1:50, Fine Biotech, China), ENNP1 (1:50, Abcam, USA), and NAT2 (1:50, Abclonal, China).

Techniques:

Journal: Frontiers in Oncology

Article Title: Mining of RNA Methylation-Related Genes and Elucidation of Their Molecular Biology in Gallbladder Carcinoma

doi: 10.3389/fonc.2021.621806

Figure Lengend Snippet: RNA methylation sites of hub genes form RMBase database.

Article Snippet: Primary antibodies against the following proteins were purchased: FGA (1:50, Boster, China), F2 (1:50, Boster, China), CFH (1:50, Boster, China), PIPOX (1:50, Absin, China), ITIH4 (1:50, Proteintech, USA), GNMT (1:50, Proteintech, USA), MAT1A (1:50, Fine Biotech, China), MTHFD1 (1:50, Proteintech, USA), HPX (1:50, Boster, China), CTH (1:50, Boster, China), CFHR3 (1:50, Fine Biotech, China), ENNP1 (1:50, Abcam, USA), and NAT2 (1:50, Abclonal, China).

Techniques: Methylation

Journal: Frontiers in Oncology

Article Title: Mining of RNA Methylation-Related Genes and Elucidation of Their Molecular Biology in Gallbladder Carcinoma

doi: 10.3389/fonc.2021.621806

Figure Lengend Snippet: Immunohistochemistry of FGA, F2, CFH, PIPOX, ITIH4, GNMT, MAT1A, MTHFD1, HPX, CTH, CFHR3, ENNP1, and NAT2 in clinical GBC and control specimens.

Article Snippet: Primary antibodies against the following proteins were purchased: FGA (1:50, Boster, China), F2 (1:50, Boster, China), CFH (1:50, Boster, China), PIPOX (1:50, Absin, China), ITIH4 (1:50, Proteintech, USA), GNMT (1:50, Proteintech, USA), MAT1A (1:50, Fine Biotech, China), MTHFD1 (1:50, Proteintech, USA), HPX (1:50, Boster, China), CTH (1:50, Boster, China), CFHR3 (1:50, Fine Biotech, China), ENNP1 (1:50, Abcam, USA), and NAT2 (1:50, Abclonal, China).

Techniques: Immunohistochemistry, Control