Journal: Biology of reproduction

Article Title: CDH1 is a specific marker for undifferentiated spermatogonia in mouse testes.

doi: 10.1095/biolreprod.106.053181

Figure Lengend Snippet: FIG. 5. Stem cell markers were expressed in a subset of CDH1-positive cells. a) Double staining of CDH1 and POU5F1. The nuclei of all CDH1-positive cells were stained with POU5F1, but the expression level of POU5F1 was low in some As spermatogonia (arrowheads). b) Double staining of CDH1 and RET. RET was expressed in almost all As and Apr cells but rarely expressed in Aal cells. c) Double staining of CDH1 and GFRA1. The staining pattern of GFRA1 was similar to that of RET. d) Double staining of CDH1 and ZBTB16. The nuclear staining of ZBTB16 was mosaic in CDH1-positive cell clusters. Nonspecific binding of the secondary antibody stained the peritubular fibroblast cells (asterisks). Bar ¼ 20 lm. e) Schematic view of the expression level of several surface antigens and POU5F1 through the early develop- ment of spermatogonia. The expression of CDH1 is reduced during the transition from undifferentiated type A spermatogonia to A1 spermatogonia. By contrast, the expression of KIT and TACSTD1 is upregulated. RET and GFRA1 are expressed mainly in As and Apr cells and expressed rarely in Aal cells.

Article Snippet: The samples were incubated with 10 lg/ml rat anti-CDH1 antibody and 10 lg/ml mouse anti-RET antibody (AF482; R&D Systems).

Techniques: Double Staining, Staining, Expressing, Binding Assay

Journal: bioRxiv

Article Title: High efficiency organoid-derived cyst cultures as a drug discovery platform for polycystic kidney disease

doi: 10.1101/2025.02.16.638545

Figure Lengend Snippet: A . Schematic of the protocol. B . Day (d) 12 nephron organoids differentiated from wildtype iPSCs. C . Enzymatic digestion of whole nephron organoids into a single cell suspension, marking d0 of UB differentiation. D . Development of UB organoids over 15 days. E . qPCR analysis comparing d12 nephron organoids and d7 UB organoids for nephron and UB marker gene expression. ** p≤ 0.01; *** p≤ 0.001; **** p≤ 0.0001 (Unpaired t -test). F . Immunohistological characterization of UB organoids showing expression of distal tubule (CDH1, PAX2, GATA3) and UB markers (RET, KRT8). Nuclear counterstain, Hoechst. Scale bars: 400 µm (B-D); 100 µm (F).

Article Snippet: Antibodies used were: PAX2 (BioLegend 901001), CDH1 (BD Biosciences 610181), RET (Novus NBP184568), GATA3 (Thermo MA1-028), KRT8 (DSHB TROMA), EPCAM (Sigma HPA026761).

Techniques: Suspension, Marker, Gene Expression, Expressing

Journal: bioRxiv

Article Title: High efficiency organoid-derived cyst cultures as a drug discovery platform for polycystic kidney disease

doi: 10.1101/2025.02.16.638545

Figure Lengend Snippet: A . Development of PKD2 -/- UB organoids over 25 days. Arrow shows an early cyst on d7. B . Quantification of cyst development in wildtype and PKD2 -/- UB organoids. C . PKD2 -/- cyst diameters increase over time. D . Example of a high efficiency cyst culture at d17. E . Upregulation of ADPKD marker genes in PKD2 -/- compared to wildtype UB organoids. ** p≤ 0.01; *** p≤ 0.001; **** p≤ 0.0001 (Unpaired t -test). F . Immunohistological analysis of PKD2 -/- UB organoids, showing expression of distal tubule and UB markers CDH1, PAX2, RET and GATA3 in the cystic epithelium. Nuclear counterstain, Hoechst. Scale bars: 400 µm (A); 1000 µm (D); 100 µm (F).

Article Snippet: Antibodies used were: PAX2 (BioLegend 901001), CDH1 (BD Biosciences 610181), RET (Novus NBP184568), GATA3 (Thermo MA1-028), KRT8 (DSHB TROMA), EPCAM (Sigma HPA026761).

Techniques: Marker, Expressing

Journal: Journal of neurochemistry

Article Title: A specific isoform of glial cell line-derived neurotrophic factor family receptor alpha 1 regulates RhoA expression and glioma cell migration.

doi: 10.1111/j.1471-4159.2010.06975.x

Figure Lengend Snippet: Fig. 2 Knockdown of GFRa1a and GFRa1b by isoform-specific siRNA. (a) Graphic representation of GFRa1a and GFRa1b exon organizations, positions of isoform-specific and total GFRa1 siRNAs. (b) Specific knockdown of GFRa1 isoforms or total GFRa1 by siRNAs and quantified by real-time PCR. Relative mRNA expressions of GFRa1a (filled bars) or GFRa1b isoform (open bars) were specifically and efficiently knocked down by two siRNAs of each iso- form (nos 1 and 2). Two siRNAs targeting common region of GFRa1 isoforms were also used to knock down the total levels of GFRa1. Luciferase siRNA was used as negative control. *p < 0.05, **p < 0.01.

Article Snippet: Small interfering RNA duplexes specific for GFRa1a and GFRa1b were designed against the unique exon junctions. siRNAs for GFRa1a and GFRa1b were from Invitrogen; siRNAs for total GFRa1 and RhoA were from Integrated DNA Technologies (IDT, Coralville, IA, USA), siRNAs for Ret (sc-156121) and NCAM (sc156119) were from Santa Cruz Biotechnologies; luciferase control siRNA was obtained from Sigma.

Techniques: Knockdown, Real-time Polymerase Chain Reaction, Luciferase, Negative Control

Journal: Journal of neurochemistry

Article Title: A specific isoform of glial cell line-derived neurotrophic factor family receptor alpha 1 regulates RhoA expression and glioma cell migration.

doi: 10.1111/j.1471-4159.2010.06975.x

Figure Lengend Snippet: Fig. 3 Knockdown of GFRa1b induced C6 cell elongation. C6 cells were transfected with control, GFRa1a, GFRa1b and total GFRa1 siRNAs and stained with fluorescein diacetate (FDA) 48 h post- transfection. (a) Representative images of C6 cells after FDA staining. White arrows indicate elongated processes of C6 cells. Scale bar, 50 lm. (b) Quantification of C6 cell elongation after GFRa1 knock- down. Similar results were obtained with two different siRNA duplexes. **p < 0.01 compared with control siRNA.

Article Snippet: Small interfering RNA duplexes specific for GFRa1a and GFRa1b were designed against the unique exon junctions. siRNAs for GFRa1a and GFRa1b were from Invitrogen; siRNAs for total GFRa1 and RhoA were from Integrated DNA Technologies (IDT, Coralville, IA, USA), siRNAs for Ret (sc-156121) and NCAM (sc156119) were from Santa Cruz Biotechnologies; luciferase control siRNA was obtained from Sigma.

Techniques: Knockdown, Transfection, Control, Staining

Journal: Journal of neurochemistry

Article Title: A specific isoform of glial cell line-derived neurotrophic factor family receptor alpha 1 regulates RhoA expression and glioma cell migration.

doi: 10.1111/j.1471-4159.2010.06975.x

Figure Lengend Snippet: Fig. 4 Knockdown of GFRa1b inhibited C6 cell migration and invasion. C6 cells were transfected with control, GFRa1a, GFRa1b or total GFRa1 siRNAs. (a) C6 cell migra- tion after siRNA transfection was examined by wound healing assay. (b) Quantification of C6 cell migration by trans-well migration assay. (c) C6 cell invasion was measured by matrigel invasion assay. Similar results were obtained with two different siRNA duplexes. **p < 0.01.

Article Snippet: Small interfering RNA duplexes specific for GFRa1a and GFRa1b were designed against the unique exon junctions. siRNAs for GFRa1a and GFRa1b were from Invitrogen; siRNAs for total GFRa1 and RhoA were from Integrated DNA Technologies (IDT, Coralville, IA, USA), siRNAs for Ret (sc-156121) and NCAM (sc156119) were from Santa Cruz Biotechnologies; luciferase control siRNA was obtained from Sigma.

Techniques: Knockdown, Migration, Transfection, Control, Wound Healing Assay, Invasion Assay