Journal: Neuroscience Insights

Article Title: Cerebral Proteomic Changes in the rTg-D Rat Model of Cerebral Amyloid Angiopathy Type-2 With Cortical Microhemorrhages and Cognitive Impairments

doi: 10.1177/26331055241288172

Figure Lengend Snippet: Arteriolar deposition of fibrillar amyloid and loss of smooth muscle cells in rTg-D rats. Brain sections from a WT rat (20 M) (A, C and E) or similarly aged rTg-D rat (22 M) (B, D and F) were immunolabeled with antibodies to collagen IV to identify cerebral blood vessels (red) or to smooth muscle cell α actin to identify smooth muscle cells (green) and stained with AmyGlo to visualize fibrillar amyloid (blue). Representative images from pial arteries (A and B); cortical arteries (C and D); and cortical arterioles (E and F). Scale bars = 50 µm.

Article Snippet: Staining for fibrillar amyloid was performed either using Amylo-Glo as described by the manufacturer (Biosensis Inc., Thebarton, South Australia) or staining with thioflavin S. The following antibodies were used for immunohistochemical analysis: rabbit polyclonal antibody to collagen type IV to visualize cerebral vessels (1:100; ThermoFisher, Rockford, IL); mouse monoclonal antibody to smooth muscle cell α actin (1:200, NBP 2-33006, Novus Bio); and antibody to HTRA1 (1:200, MAB2916, R&D Systems).

Techniques: Immunolabeling, Staining

Journal: Scientific Reports

Article Title: Sustained alterations in proximal tubule gene expression in primary culture associate with HNF4A loss

doi: 10.1038/s41598-024-73861-3

Figure Lengend Snippet: HNF4A activity is central to expression changes in culture. ( a ) ChEA3 transcription factor network map of genes downregulated in the first day after plating. Full results are available in Supplementary Data . ( b ) Heatmap of proximal tubule HNF4A targets in cultured cells over time. Row Z-scores are shown and are available in Supplementary Data . ( c ) Expression of Hnf4a and two of its proximal tubule targets, Slc34a1 and Pck1 . Confirmatory quantitative RT-PCR from independent samples are shown in Supplementary Fig . ( d ) Gene Set Enrichment Analysis (GSEA) plots for HNF4A target genes defined by Marable et al. in the differentially expressed genes after 1 day (vs. 0 day) and 14 days (vs. 0 day) of culture.

Article Snippet: A retrovirus vector was generated using an HNF4A overexpression (HNF4A-OE) vector (Addgene plasmid #33006).

Techniques: Activity Assay, Expressing, Cell Culture, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Sustained alterations in proximal tubule gene expression in primary culture associate with HNF4A loss

doi: 10.1038/s41598-024-73861-3

Figure Lengend Snippet: De-differentiation is insensitive to culture conditions. ( a ) Legend and details for specific culture condition changes. ( b ) Quantitative RT-PCR for Hnf4a and Havcr1 in fresh enriched proximal tubules (TUBULES) vs. 14 days cultures (* P ≤ 0.05, ANOVA with Tukey post hoc test relative to standard conditions (STD) at 14 days; all samples were highly significant compared with tubules, N = 4–9).

Article Snippet: A retrovirus vector was generated using an HNF4A overexpression (HNF4A-OE) vector (Addgene plasmid #33006).

Techniques: Quantitative RT-PCR