Journal: Communications Medicine

Article Title: Transplantation of decellularized porcine kidney grafts repopulated with primary human cells demonstrates filtration function in pigs

doi: 10.1038/s43856-024-00676-8

Figure Lengend Snippet: a – c Representative immunofluorescence images comparing cultured glomerular outgrowth cells (GOCs) pre-differentiation, and d – f 6 days post-differentiation. a , d Podocyte differentiation is depicted through changes in F-actin organization (green). b , e Expression of podocyte lineage marker podocin (red). c , f Expression of podocyte lineage marker podocalyxin (green). 4′,6-diamidino-2-phenylindole (DAPI) was used to label nuclei blue. g RT-qPCR was performed to assess relative expression of podocyte marker genes synaptopodin ( SYNPO ) and nephrin ( NPHS1 ), comparing mRNA levels of GOCs from 3 separate donor lots cultured for 6 days in control, endothelial growth media (EGM) compared to PSM. Relative to control media, PSM induced a 7-fold and 195-fold increase in SYNPO and NPHS1 mRNA, respectively. Data is represented as mean ± SD of three independent biological replicates ( n = 3). Statistical difference in target genes between EGM control media and PSM podocyte differentiation media was determined with one-tailed t-test: * p = 0.0346, ** p = 0.002. Source data are provided in Supplementary Data .

Article Snippet: TaqMan Probe IDs: Nephrin Hs00190446_m1 NPHS1 , Synaptopodin Hs00200768_m1 SYNPO , HMBS Hs00609297_m1 HMBS .

Techniques: Immunofluorescence, Cell Culture, Expressing, Marker, Quantitative RT-PCR, Control, One-tailed Test

Journal: Frontiers in Pharmacology

Article Title: Sanqi Oral Solution Mitigates Proteinuria in Rat Passive Heymann Nephritis and Blocks Podocyte Apoptosis via Nrf2/HO-1 Pathway

doi: 10.3389/fphar.2021.727874

Figure Lengend Snippet: SQ alleviated glomerular podocyte injury in PHN rats. Effects of SQ and CP on foot process width (magnification × 12,000, red arrows) and synaptopodin expression (magnification × 400) were measured by TEM and immunofluorescence staining (A) . With the treatment of SQ and CP, restored glomerular podocytic foot processes (B) and synaptopodin expression (C) were seen in PHN rats ( n = 6). Data are represented as mean ± SD from independent groups. ** p < 0.01 vs. Control group. ## p < 0.01 vs. Model group.

Article Snippet: Primary antibodies against C5b-9 (sc-66190) and synaptopodin (sc-515842) were purchased from Santa Cruz Biotechnology (Dallas, Texas, United States).

Techniques: Expressing, Immunofluorescence, Staining, Control

Journal: American Journal of Translational Research

Article Title: Distinct effects of ANGPT2 on gene expression of glomerular podocytes and mesangial cells

doi:

Figure Lengend Snippet: ANGPT2 protein expression in glomerular cells of human and mouse. A. Immunofluorescence staining of ANGPT2 in normal human glomeruli. Podocyte marker, SYNPO, was co-stained, thereby localizing ANGPT2 to podocytes and non-podocytes. B. Similar co-staining of ANGPT2 and SYNPO in mouse glomeruli showing ANGPT2 is present in both podocytes and non-podocytes in mice.

Article Snippet: The information of the reagents used in the present study is as follows: Antibodies against ANGPT2 (Affinity), SYNPO (Santa Cruz), GAPDH (Proteintech), total ERK1/2, phospho-ERK1/2, total AKT and phospho-AKT (Cell Signaling Technologies); Reverse transcription kit (DRR037A, Takara); Quantitative PCR kit (Thermo Fisher Scientific); RIPA cell lysis buffer, BCA protein quantification kit (Beyotime, Shanghai); RNA extraction kit (Takara); ANGPT2 (HY-P7510, MCE); Trizol (Invitrogen).

Techniques: Expressing, Immunofluorescence, Staining, Marker

Journal: Cellular and molecular life sciences : CMLS

Article Title: Deficiency of the Src homology phosphatase 2 in podocytes is associated with renoprotective effects in mice under hyperglycemia

doi: 10.1007/s00018-022-04517-6

Figure Lengend Snippet: Upregulation of Shp2 in the kidney and E11 podocyte cell line under hyperglycemia. a,b Immunoblots of Shp2 and Actin (a), and Ptpn11 mRNA expression (b) in kidneys from male mice fed with regular chow (Ctrl), HFD (24 weeks), or challenged with STZ (160 μg/g body weight, 20 weeks). Actin served as a protein loading control for immunoblotting (n=4 per group), and mRNA expression was normalized to Tbp (n=4 per group). Each lane in immunoblot represents a sample from an individual mouse. *p ≤ 0.05 indicates a significant difference between Ctrl versus STZ-treated and HFD-fed mice. c Confocal images of kidney sections from Ctrl, HFD-fed (24 weeks), and STZ-treated (160 μg/g body weight, 20 weeks) wild-type male mice co-immunostained for Shp2 (green) and Synaptopodin (Synpo, red). Scale bar: 20 μm. The boxed areas in the Merge are enlarged in the lower panel. d Differentiated E11 podocytes were treated with high glucose (25 mM; HG) for 0, 24, 48, and 72 hours. Cell lysates were immunoblotted for Shp2, Synpo, and Actin. The protein expression level was normalized to Actin from three independent experiments. Statistical significance in (d) was considered *p ≤ 0.05 for Shp2 and # p ≤ 0.05 for Synpo between 0h and 24h, 48h, and 72 h. A.U.: arbitrary unit.

Article Snippet: Sections were stained using antibodies for Shp2 (sc-280), Nephrin (sc-19000) (both from Santa Cruz Biotechnology), Synaptopodin (Novus Biologicals; NBP2–33357), pAKT Ser473 (4060), pERK1/2 Tyr202/Thr204 (4370) (both from Cell Signaling Technology), and Collagen III (Abcam; ab7778) at 4°C overnight.

Techniques: Western Blot, Expressing, Control

Journal: Kidney360

Article Title: Decay-Accelerating Factor Expression Modulates the Severity of Experimental Focal Segmental Glomerulosclerosis

doi: 10.34067/kid.0005312022

Figure Lengend Snippet: Figure 2. Pharmacologic inhibition of DAF enzymatic cleavage prevents ADR-induced FSGS in WT BALB/c mice. (A) Representative pictures and (B) data quantification of glomerular expression of DAF and synaptopodin in WT BALB/c mice treated with vehicle control (DMSO) (n54) or FIPI (n54) starting from the day of ADR injection. (C) Representative pictures and (D) data quantification of the glomerular expression of C3b and synaptopodin in WT BALB/c mice treated with vehicle control or FIPI starting from the day of ADR injection. Scale bars: 50 mm. (E) Urinary albumin-to-creatinine ratio (ACR) over time and (F) representative light microscopy images (PAS, scale bars: 20 mm) of glomeruli in the same mice. Statistical analysis was performed using unpaired t test for immunofluorescence and two-way ANOVA with ˇSida´k test for multiple comparisons for ACR. *P,0.05; **P,0.01; ***P,0.001; ****P,0.0001.

Article Snippet: Immunofluorescence staining was performed on cryosections (5 mm thick) as per our protocol9 using the following primary antibodies: anti-DAF antibody (rabbit anti-mouse, 1:50; Thermo Fisher), anti-C3b antibody (rat anti-mouse, 1: 50; Hycult Biotech, Wayne, PA), and synaptopodin (antimouse, 1:5; Fitzgerald, Acton, MA) at 4°C overnight.

Techniques: Inhibition, Expressing, Control, Injection, Light Microscopy