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: Fig. 4. HNF4A activity is central to expression changes in culture. (a) ChEA339 transcription factor network map of genes downregulated in the first day after plating. Full results are available in Supplementary Data S3. (b) Heatmap of proximal tubule HNF4A targets42 in cultured cells over time. Row Z-scores are shown and are available in Supplementary Data S2. (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 S7. (d) Gene Set Enrichment Analysis (GSEA) plots for HNF4A target genes defined by Marable et al.42 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) vector44 (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: Fig. 5. 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) vector44 (Addgene plasmid #33006).

Techniques: 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: 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