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: Molecular and cellular endocrinology

Article Title: SDHB and SDHD silenced pheochromocytoma spheroids respond differently to tumour microenvironment and their aggressiveness is inhibited by impairing stroma metabolism.

doi: 10.1016/j.mce.2022.111594

Figure Lengend Snippet: Fig. 1. Characterization of stable SDHB and SDHD silenced cell lines. (A) Representative blots of the expression of mitochondrial SDHA, B and D subunits. (B) Densitometric analysis of western blot bands, performed by Bio-Rad imaging and analysis software (Quantity One), showed significant differences in the SDHB and SDHD subunit expression levels in SDHB (light grey) and in SDHD silenced cells (dark grey) respectively, compared to Wt (black). Bars are the means of three independent preparations ± SD, ***p < 0.001. (C) Representative traces of SDH enzymatic activity measured in cell homogenates. The silenced SDHB and SDHD cells (dotted and continuous lines, respectively) showed a similar decrease of the SDH activity, significantly different compared with Wt (dashed line). (D) Histogram represents the SDH activity expressed as the percentages. SDHB and SDHD silenced cells (light and dark grey, respectively) showed a significantly decreased of SDH activity compared to Wt (black). Bars are the means of three independent experiments (each of them conducted in duplicate samples) ± SD, ***p < 0.01. (E) The bar graph represents the means of intracellular succinate/fumarate ratio ± SD, measured by GC/MS, in three independent experiments with two replicates. SDHB silenced cells (light grey) showed a significant increase of the metabolites ratio compared with both Wt (black) and to SDHD silenced cells (dark grey), *p < 0.05, **p < 0.01. (F) Representative immunoblot of HIF1α expression in Wt, SDHB and SDHD silenced cells. (G) Optical density analysis of western blot bands. Actin im munoblots was used as loading control. For all the analyses One-way ANOVA post-test Bonferroni was used.

Article Snippet: Bound antibodies were detected using ECL reagents (Immobilon) and analysed with a Bio-Rad ChemiDoc Imaging System (Quantity One) for dedicated chemiluminescent image acquisition.

Techniques: Expressing, Western Blot, Imaging, Software, Activity Assay, Gas Chromatography-Mass Spectrometry, Control