Journal: PLoS ONE

Article Title: Lack of APOL1 in proximal tubules of normal human kidneys and proteinuric APOL1 transgenic mouse kidneys

doi: 10.1371/journal.pone.0253197

Figure Lengend Snippet: A . Immunofluorescent staining for APOL1, nephrin (to identify podocytes), and CD31 (to identify endothelia cells) in the BAC-APOL1 transgenic mouse kidney (G1 mouse is shown). B. Comparison of APOL1 expression patterns in all three APOL1 transgenic lines. Proximal tubules were identified by labeling with fluorescent Lotus tetragonolobus (LT) lectin. APOL1 was present in vascular endothelia (arrow heads), in podocytes (arrows), and trapped in vascular spaces (*), but not tubular epithelia. Scale bar = 40μm.

Article Snippet: FITC-labeled Lotus tetragonolobus lectin (Vector labs) was used to label proximal tubule cells as previously described [ ].

Techniques: Staining, Transgenic Assay, Expressing, Labeling

Journal: PLoS ONE

Article Title: Lack of APOL1 in proximal tubules of normal human kidneys and proteinuric APOL1 transgenic mouse kidneys

doi: 10.1371/journal.pone.0253197

Figure Lengend Snippet: A . Control immunostaining in non-transgenic (wildtype) and proteinuric HIVAN4 mice for APOL1 along with fluorescent Lotus tetragonolobus (“LT”) lectin binding to demarcate the proximal tubule brush border. Since wildtype and HIVAN4 mice do not have APOL1, the staining observed in parietal cells (arrows) is artifact. B . Immunostaining for APOL1 in proteinuric BAC transgenic mice of each APOL1 genotype (representative images are shown, number of animal examined in each genotype group were the same as for ). Images show proximal tubules at the transition with Bowman capsule. The boxed region is magnified below each panel along with the isolated fluorescent channels shown in black and white. C . Positive control immunostaining for a filtered lipoprotein, APOA1, and fluorescently-labelled Lotus tetragonolobus (“LT”) lectin. An APOL1-G0 mouse and an APOL1-G0 x HIVAN4 dual transgenic mouse with proteinuria is shown; below each respective color panel is the individual fluorescent channels (in black and white) of the boxed region for either LT lectin or APOA1. White arrows mark glomerular capillaries containing circulating APOA1 protein within capillary lumens, red arrows denote APOA1 in protein reabsorption droplets at the brush border of proximal tubules. Scale bar = 40μm.

Article Snippet: FITC-labeled Lotus tetragonolobus lectin (Vector labs) was used to label proximal tubule cells as previously described [ ].

Techniques: Immunostaining, Transgenic Assay, Binding Assay, Staining, Isolation, Positive Control

Journal: Aging and disease

Article Title: FUT8-Mediated Core Fucosylation Promotes the Pulmonary Vascular Remodeling in Pulmonary Arterial Hypertension

doi: 10.14336/ad.2023.0218

Figure Lengend Snippet: Figure 3. 2FF improved the hemodynamics and pulmonary vascular remodeling in MCT-induced PAH rats. Weight changes of rats in control (N=4), MCT and 2FF treatment groups (N=6) are shown in (A), and RVSP of rats detected by right heart catheterization is shown in (B). Right heart hypertrophy index (C) and right heart weight ratio (D) of rats in the three groups were measured. Immunofluorescence (E) and HE staining (F) were used to evaluate the arterial media thickness and the arrow indicates the pulmonary arterioles. Scale bar=50 μm. (G) Protein expression levels of FUT8, PCNA, Bcl-2/Bax in control (n=4), MCT and 2FF treatment groups (n=6) were measured using western blotting. β-actin was used as a housekeeper protein for normalization. Quantifications of the protein levels are shown. (H) Core fucosylation levels of homogenous proteins in control (n=4), MCT and 2FF treatment-rat (n=6) lung tissues were measured using lectin blotting, and β-actin was used as a housekeeper protein for normalization. Quantification of core fucosylation level is shown. All results are shown as mean ± SD. Data were analyzed by one way ANOVA. MCT group was compared with the control group and 2FF treatment group separately. p-values are indicated above the groups being compared with each other. Ns indicates no statistically significant.

Article Snippet: A Human FUT8 ELISA Kit (EK1990, Boster Biological Technology Company) and an ELISA Kit for rats (FY40556, FeiYa Biotechnology) were used to detect the concentration of FUT8 according to the manufacturer’s instructions.

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

Journal: Aging and disease

Article Title: FUT8-Mediated Core Fucosylation Promotes the Pulmonary Vascular Remodeling in Pulmonary Arterial Hypertension

doi: 10.14336/ad.2023.0218

Figure Lengend Snippet: Figure 4. FUT8 was upregulated in both PAH patients and our rat experimental model. Concentrations of FUT8 in human (A) and rat (B-C) blood samples were detected by ELISA. There were 11 volunteers, 11 patients with CTD- PAH or IPAH, and 13 patients with CHD-PAH (diagnosed PAH by right heart catheterization in Xiangya Hospital). The blood samlpes of rats were from 8 control rats and 11 rats injected MCT. (D) Immunofluorescence of α-SMA and FUT8 in rat lung tissue slice is shown, Scale bar=50 μm. (E) Protein expression levels of FUT8 and PCNA in control and MCT-rat lung tissue homogenate were measured using western blotting (n=4). β-actin was used as a housekeeper protein for normalization. Quantifications of the protein levels are shown. (F) mRNA exprssion of FUT8 in rat lung tissue homogenization was determined by qPCR (n=4). (G) Protein expression levels of FUT8 (n=4) and PCNA (n=5) in control and PDGF-PASMCs were measured useing western blotting. (H) Immunofluorescence of FUT8 in control and PDGF-PASMCs is shown. All results are shown as mean ± SD. Data of three groups in (A) were analyzed by one way ANOVA, and the Mann-Whitney U test was performed for the comparisons between control and MCT (or PDGF) groups in other charts. p-values are indicated above the groups being compared with each other.

Article Snippet: A Human FUT8 ELISA Kit (EK1990, Boster Biological Technology Company) and an ELISA Kit for rats (FY40556, FeiYa Biotechnology) were used to detect the concentration of FUT8 according to the manufacturer’s instructions.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Injection, Immunofluorescence, Expressing, Western Blot, Tissue Homogenization, MANN-WHITNEY

Journal: Aging and disease

Article Title: FUT8-Mediated Core Fucosylation Promotes the Pulmonary Vascular Remodeling in Pulmonary Arterial Hypertension

doi: 10.14336/ad.2023.0218

Figure Lengend Snippet: Figure 5. FUT8 silencing impairs proliferation, migration, apoptosis resistance, and phenotype conversion of PASMCs. (A) Proliferative activity of PASMCs treated with PDGF or siRNA was detected using the CCK8 assay (n=6). (B) Protein expression levels of FUT8 (n=4), PCNA (n=4), Bcl-2/Bax (n=4), OPN (n=5) in control and PDGF- PASMCs treated with or without knocking down FUT8 were measured using Western blotting. β-actin was used as a housekeeper protein for normalization. Quantifications of protein levels are shown. (C) Apoptotic rate of PASMCs- knocked down FUT8 was detemined using flow cytometry (n=5). (D) Effect of knocking down FUT8 on migration capacity of PASMCs was determined using the scratch experiment (n=4), Scale bar=100 μm. All results are shown as mean ± SD. Data were analyzed by one way ANOVA. PDGF+siNC vs. control+siNC group; PDGF+siFUT8 vs. PDGF+siNC group. p-values are indicated above the groups being compared with each other.

Article Snippet: A Human FUT8 ELISA Kit (EK1990, Boster Biological Technology Company) and an ELISA Kit for rats (FY40556, FeiYa Biotechnology) were used to detect the concentration of FUT8 according to the manufacturer’s instructions.

Techniques: Migration, Activity Assay, CCK-8 Assay, Expressing, Control, Western Blot, Flow Cytometry

Journal: Aging and disease

Article Title: FUT8-Mediated Core Fucosylation Promotes the Pulmonary Vascular Remodeling in Pulmonary Arterial Hypertension

doi: 10.14336/ad.2023.0218

Figure Lengend Snippet: Figure 7. Model diagram describing the effect of FUT8 and its regulated core fucosylation on PASMCs. FUT8 expression may be increased by the stimulation of hypoxia, inflammation, or growth factors in PASMCs. Core fucosylation of whole cell proteins, including VEGFR2, is increased, which promotes activation of the AKT pathway, thereby promoting cell proliferation and migration, apoptotic resistance, and transformation to a synthetic phenotype, ultimately facilitating the process of vascular remodeling in PAH.

Article Snippet: A Human FUT8 ELISA Kit (EK1990, Boster Biological Technology Company) and an ELISA Kit for rats (FY40556, FeiYa Biotechnology) were used to detect the concentration of FUT8 according to the manufacturer’s instructions.

Techniques: Expressing, Activation Assay, Migration, Transformation Assay