amsbio ams a2506  (AMS Biotechnology)

Journal: Journal of nanobiotechnology

Article Title: Ceria-Zirconia nanoparticles reduce intracellular globotriaosylceramide accumulation and attenuate kidney injury by enhancing the autophagy flux in cellular and animal models of Fabry disease.

doi: 10.1186/s12951-022-01318-8

Figure Lengend Snippet: Fig. 3 Gb3 detection in α-GLA knockdown HK-2 cells and human podocytes at baseline and under PEG-CZNPs treatment. A–D HK-2 cell analysis A Representative confocal immunofluorescence microscopy images of Gb3 (green), with DAPI counterstaining (blue), in control (siRNA) and α-GLA deficient HK-2 cells (siGLA), with or without exposure to PEG-CZNPs. B Quantification of the fluorescence intensities in the confocal microscope images using ImageJ software. C Representative TEM images of control (siRNA) and α-GLA deficient HK-2 cells (siGLA), with or without exposure to PEG-CZNPs. D Quantitative analysis of Gb3 vesicles in control (siRNA) and α-GLA deficient HK-2 cells (siGLA), with or without exposure to PEG-CZNPs. E–I Human podocyte analysis E Representative confocal immunofluorescence microscopy images of Gb3 (green), with DAPI counterstaining (blue), in control (siRNA) and α-GLA deficient human podocytes (siGLA) with or without exposure to PEG-CZNPs. F Quantification of the fluorescence intensity of confocal microscope images using ImageJ software. G Representative TEM images of control (siRNA) and α-GLA deficient human podocytes (siGLA) with or without exposure to PEG-CZNPs. H Quantitative analysis of Gb3 vesicles from TEM images. I Quantification of the total Gb3 content using by LC–MS/MS data and the Student’s t test. Data shown are the mean ± SD; *P < 0.05, **P < 0.01, and ***P < 0.001 versus control; #P < 0.05, ##P < 0.01, and ###P < 0.001 versus siGLA knockdown alone

Article Snippet: IFA was also performed Using primary antibodies against Gb3 (Amsbio, Cambridge, MA), synaptopodin (Sigma Aldrich) and LC3B (Sigma Aldrich).

Techniques: Knockdown, Cell Analysis, Immunofluorescence, Microscopy, Control, Fluorescence, Software, Liquid Chromatography with Mass Spectroscopy

Journal: Journal of nanobiotechnology

Article Title: Ceria-Zirconia nanoparticles reduce intracellular globotriaosylceramide accumulation and attenuate kidney injury by enhancing the autophagy flux in cellular and animal models of Fabry disease.

doi: 10.1186/s12951-022-01318-8

Figure Lengend Snippet: Fig. 12 Schematic illustrations of the PEG-CZNP induced attenuation of kidney injury from FD. This occurs via the enhancement of the autophagy flux combined with the activation of TFEB, restoration of AKT/mTOR signaling, and antioxidant effects. PEG-CZNPs thereby alleviate inflammatory and fibrosis pathways and attenuate Gb3-mediated kidney injury

Article Snippet: IFA was also performed Using primary antibodies against Gb3 (Amsbio, Cambridge, MA), synaptopodin (Sigma Aldrich) and LC3B (Sigma Aldrich).

Techniques: Activation Assay

Journal: Experimental & Molecular Medicine

Article Title: Human kidney organoids reveal the role of glutathione in Fabry disease

doi: 10.1038/s12276-021-00683-y

Figure Lengend Snippet: a Representative image of immunofluorescence staining of Gb3, LTL and NPHS1 in WT and GLA-mutant kidney organoids. Scale bar = 50 µm. b Quantification of the percentage of Gb3-positive area. c Representative images of immunofluorescence staining of NPHS1 and Gb3 overlaid with the corresponding immunogold TEM images labeled with Gb3 obtained from the same field in GLA-mutant kidney organoids. Scale bar = 100 µm for ( c ), 25 µm for ( c ) Panel ( a ), 2 µm for ( c ) Panel ( b ), 1 µm for ( c ) Panel ( c ), and 0.5 µm for ( c ) Panel ( d ). d Representative oil red O staining images showing lipid droplet formation in podocytes and tubules. Scale bar = 30 µm, 10 µm for zoom. e Representative TEM images showing lipid droplets (red arrowhead). Scale bar = 2 µm 1 µm for e panel. f Quantification of the number of lipid droplets. The values are the mean ± SEM. * p < 0.05; ** p < 0.01.

Article Snippet: The sections were labeled at 4 °C overnight using a mouse polyclonal antibody against Gb3 (TCI chemicals A2506, 1:300 dilution).

Techniques: Immunofluorescence, Staining, Mutagenesis, Labeling

Journal: Experimental & Molecular Medicine

Article Title: Human kidney organoids reveal the role of glutathione in Fabry disease

doi: 10.1038/s12276-021-00683-y

Figure Lengend Snippet: a Representative brightfield image of changes in the morphology of GLA-mutant kidney organoids after ERT. Scale bar = 300 µm. b Quantification of kidney organoid diameters after ERT. c Representative western blot of the expression of GLA after ERT. d Representative image of immunofluorescence staining of Gb3 in GLA-mutant kidney organoids after ERT. Scale bar = 50 µm. e Quantification of the percentage of Gb3-positive area. The values are the mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The sections were labeled at 4 °C overnight using a mouse polyclonal antibody against Gb3 (TCI chemicals A2506, 1:300 dilution).

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

Journal: Experimental & Molecular Medicine

Article Title: Human kidney organoids reveal the role of glutathione in Fabry disease

doi: 10.1038/s12276-021-00683-y

Figure Lengend Snippet: a Immunofluorescence staining of globotriaosylceramide (Gb3) and markers of podocytes (NPHS1) and proximal tubules (LTL) in GLA-mutant kidney organoids and GSH-treated GLA-mutant kidney organoids. Scale bar = 50 µm. b qRT–PCR analysis of NPHS1, PODXL, ECAD, and CDH16. c TUNEL staining images showing apoptotic cells. Scale bar = 50 µm. d Quantification of the percentage of apoptotic cells. The values are the mean ± SEM. * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: The sections were labeled at 4 °C overnight using a mouse polyclonal antibody against Gb3 (TCI chemicals A2506, 1:300 dilution).

Techniques: Immunofluorescence, Staining, Mutagenesis, Quantitative RT-PCR, TUNEL Assay