anti nephrin  (Bioss)

Journal: Renal Failure

Article Title: Aucubin ameliorates diabetic kidney disease by restoring hGENCs autophagy through promoting phosphorylation of ATG4B protein

doi: 10.1080/0886022X.2025.2605756

Figure Lengend Snippet: Protective effect of aucubin on DKD mice by promoting ATG4B protein phosphorylation and increasing autophagy. The expression levels of p-ATG4B proteins, ATG4B proteins, autophagy protein (LC3, p62, ATG5, and ATG7), apoptosis-related proteins (Bax, cleaved caspase3, and Bcl-2), EndMT marker proteins (CD31 and α-SMA), and nephrin protein in each group were detected by WB and the statistical analysis result of the proteins expression levels. *p < .05; ** p < .01. DKD: diabetic kidney disease; AU: aucubin.

Article Snippet: Proteins were transferred to polyvinylidene difluoride membranes (PVDF) and then incubated with specific primary antibodies against ATG4B, p-ATG4B(Ser383), ATG5, ATG7 (1:1,000, CST); Bcl-2, Bax (1:500, Immunoway); p62, cleaved caspase3 (1:500, Immunoway); LC3 (1:1,000, Abcam); CD31, Vimentin (1:2,000, Proteintech); nephrin (1:1,000, Affinity) and α-SMA, GAPDH(1:1,000, Proteintech), and secondary antibodies conjugated to HRP (Proteintech).

Techniques: Phospho-proteomics, Expressing, Marker

Journal: RNA Biology

Article Title: EIF4A3-induced circFAT1 promotes high glucose-induced podocyte damage via miR-30e-5p/SOX4 axis

doi: 10.1080/15476286.2025.2563865

Figure Lengend Snippet: Elevated expression of circFAT1 in HG-treated HPCs. (A) cell morphology (left) and viability measured by CCK-8 assay (right) after 48 h treatments. (B) immunofluorescence staining of Nephrin and FSP1. (C) CircFAT1 expression by qRT-PCR. (D) schematic diagram of the composition of circFAT1. (E) PCR-agarose gel electrophoresis was used to verify the back splicing sequence and RT-PCR product of circFAT1. (F) the circFAT1 and FAT1 mRNA abundance after RNase R treatment was determined via qRT-PCR. (G) RNA stability after actinomycin D (5 μg/mL) treatment. (H) nuclear and cytoplasmic fractionation showed the distribution of circFAT1 in HPCs. *** p < 0.001.

Article Snippet: The following antibodies were utilized: WT-1 antibody (#12609–1-AP, 1: 1000), Nephrin antibody (#66970–1-Ig, 1:1000), FSP1 antibody (#20886–1-AP, 1:5000) and Desmin antibody (#67793–1-Ig, 1:5000) and GAPDH antibody (#60004–1-Ig 1:10000) were obtained from Proteintech (Wuhan, China).

Techniques: Expressing, CCK-8 Assay, Immunofluorescence, Staining, Quantitative RT-PCR, Agarose Gel Electrophoresis, Sequencing, Reverse Transcription Polymerase Chain Reaction, Fractionation

Journal: RNA Biology

Article Title: EIF4A3-induced circFAT1 promotes high glucose-induced podocyte damage via miR-30e-5p/SOX4 axis

doi: 10.1080/15476286.2025.2563865

Figure Lengend Snippet: Depletion of circFAT1 attenuates HG-induced podocyte migration and EMT. (A) levels of circFAT1 and FAT1 in HPCs transfected with si-circFAT1. (B) the apoptosis of HPCs was determined using flow cytometry. (C) the motility of HPCs was determined using Transwell assays. (D) Western blotting demonstrated the expression of WT-1, Nephrin, FSP1, and Desmin under different conditions. (E) immunofluorescence staining of Nephrin and FSP1. Bar = 50 μm, * p < 0.05, *** p < 0.001.

Article Snippet: The following antibodies were utilized: WT-1 antibody (#12609–1-AP, 1: 1000), Nephrin antibody (#66970–1-Ig, 1:1000), FSP1 antibody (#20886–1-AP, 1:5000) and Desmin antibody (#67793–1-Ig, 1:5000) and GAPDH antibody (#60004–1-Ig 1:10000) were obtained from Proteintech (Wuhan, China).

Techniques: Migration, Transfection, Flow Cytometry, Western Blot, Expressing, Immunofluorescence, Staining

Journal: RNA Biology

Article Title: EIF4A3-induced circFAT1 promotes high glucose-induced podocyte damage via miR-30e-5p/SOX4 axis

doi: 10.1080/15476286.2025.2563865

Figure Lengend Snippet: SOX4 overexpression attenuated the protective effect of circFAT1 knockdown against HG-induced podocyte damage. (A) SOX4 mRNA expression in HPCs transfected with SOX4-expressing plasmids. *** p < 0.001. (B) SOX4 protein expression in HPCs transfected with SOX4-expressing plasmids. *** p < 0.001 (C) SOX4 protein expression in HPCs transfected with NC, si-circFAT1, si-circFAT1 + Vector or si-circFAT1 + SOX4_oe, under HG condition. (D) apoptosis by annexin V/PI staining. (E) migration by Transwell assay. (F) protein levels of podocyte (WT-1, Nephrin) and mesenchymal (FSP1, Desmin) markers. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The following antibodies were utilized: WT-1 antibody (#12609–1-AP, 1: 1000), Nephrin antibody (#66970–1-Ig, 1:1000), FSP1 antibody (#20886–1-AP, 1:5000) and Desmin antibody (#67793–1-Ig, 1:5000) and GAPDH antibody (#60004–1-Ig 1:10000) were obtained from Proteintech (Wuhan, China).

Techniques: Over Expression, Knockdown, Expressing, Transfection, Plasmid Preparation, Staining, Migration, Transwell Assay