Journal: The American Journal of Pathology

Article Title: Injured Proximal Tubular Epithelial Cells Lose Hepatocyte Nuclear Factor 4α Expression Crucial for Brush Border Formation and Transport

doi: 10.1016/j.ajpath.2025.01.011

Figure Lengend Snippet: Primary proximal tubular epithelial cells (PTECs) lose expression of proximal tubular markers during culture. PTECs harvested directly following tissue dissociation, preseeding (P0), and at consecutive passages (P1 to P4), followed by immunohistochemical staining, show a rapid reduction of hepatocyte nuclear factor 4α (HNF4A) expression. Expression of the proximal tubular marker CD10 is also down-regulated, but this is more protracted in time. In contrast, the scattered tubular cell marker vimentin (VIM) is rapidly and highly induced in cultured PTECs. n = 3. Scale bars = 100 μm.

Article Snippet: Renal PTECs (RPTECs), purchased from Lonza, harvested at passage 2 to 5 (P2 to P5), and subjected to immunohistochemical staining, show an expression pattern consistent with that in the current cultured primary PTECs [ie, reduced expression of hepatocyte nuclear factor 4α (HNF4A) and CD10, and high vimentin (VIM) expression].

Techniques: Expressing, Immunohistochemical staining, Staining, Marker, Cell Culture

Journal: The American Journal of Pathology

Article Title: Injured Proximal Tubular Epithelial Cells Lose Hepatocyte Nuclear Factor 4α Expression Crucial for Brush Border Formation and Transport

doi: 10.1016/j.ajpath.2025.01.011

Figure Lengend Snippet: The proximal tubular phenotype is partially restored in primary proximal tubular epithelial cells (PTECs) after hepatocyte nuclear factor 4α (HNF4A) transduction. A: Gene Set Enrichment Analysis (GSEA) of differentially expressed genes (DEGs) following HNF4A transduction of PTECs using Gene Ontology biological process displays overrepresentation of gene sets associated with transport and absorption. B: GSEA using Gene Ontology cellular component (GO-CC) shows overrepresentation of gene sets linked to brush border. C and D: Enrichment plots of GO-CC gene sets apical part of cell and brush border, respectively, demonstrate significant enrichment. E and F: Gene sets of proximal tubule S1 to S2 and S3 segments, respectively, from single-nucleus RNA sequencing of adult human kidney were shown to be significantly enriched among DEGs in HNF4A-transduced cells (figures from our data analysis). False discovery rate (FDR) < 0.25 is considered statistically significant. NES, normalized enrichment score.

Article Snippet: Renal PTECs (RPTECs), purchased from Lonza, harvested at passage 2 to 5 (P2 to P5), and subjected to immunohistochemical staining, show an expression pattern consistent with that in the current cultured primary PTECs [ie, reduced expression of hepatocyte nuclear factor 4α (HNF4A) and CD10, and high vimentin (VIM) expression].

Techniques: Transduction, RNA Sequencing

Journal: The American Journal of Pathology

Article Title: Injured Proximal Tubular Epithelial Cells Lose Hepatocyte Nuclear Factor 4α Expression Crucial for Brush Border Formation and Transport

doi: 10.1016/j.ajpath.2025.01.011

Figure Lengend Snippet: Primary proximal tubular epithelial cells (PTECs) lose expression of proximal tubular markers during culture. PTECs harvested directly following tissue dissociation, preseeding (P0), and at consecutive passages (P1 to P4), followed by immunohistochemical staining, show a rapid reduction of hepatocyte nuclear factor 4α (HNF4A) expression. Expression of the proximal tubular marker CD10 is also down-regulated, but this is more protracted in time. In contrast, the scattered tubular cell marker vimentin (VIM) is rapidly and highly induced in cultured PTECs. n = 3. Scale bars = 100 μm.

Article Snippet: Purchased renal PTECs from Lonza exhibited a staining pattern that matched serially passaged PTECs, and a negligible fraction of renal PTECs displayed HNF4A positivity ( ).

Techniques: Expressing, Immunohistochemical staining, Staining, Marker, Cell Culture

Journal: The American Journal of Pathology

Article Title: Injured Proximal Tubular Epithelial Cells Lose Hepatocyte Nuclear Factor 4α Expression Crucial for Brush Border Formation and Transport

doi: 10.1016/j.ajpath.2025.01.011

Figure Lengend Snippet: The proximal tubular phenotype is partially restored in primary proximal tubular epithelial cells (PTECs) after hepatocyte nuclear factor 4α (HNF4A) transduction. A: Gene Set Enrichment Analysis (GSEA) of differentially expressed genes (DEGs) following HNF4A transduction of PTECs using Gene Ontology biological process displays overrepresentation of gene sets associated with transport and absorption. B: GSEA using Gene Ontology cellular component (GO-CC) shows overrepresentation of gene sets linked to brush border. C and D: Enrichment plots of GO-CC gene sets apical part of cell and brush border, respectively, demonstrate significant enrichment. E and F: Gene sets of proximal tubule S1 to S2 and S3 segments, respectively, from single-nucleus RNA sequencing of adult human kidney were shown to be significantly enriched among DEGs in HNF4A-transduced cells (figures from our data analysis). False discovery rate (FDR) < 0.25 is considered statistically significant. NES, normalized enrichment score.

Article Snippet: Purchased renal PTECs from Lonza exhibited a staining pattern that matched serially passaged PTECs, and a negligible fraction of renal PTECs displayed HNF4A positivity ( ).

Techniques: Transduction, RNA Sequencing