Journal: Nature Communications

Article Title: FGF4-FGFR1 signaling promotes podocyte survival and glomerular function to ameliorate diabetic kidney disease in male mice

doi: 10.1038/s41467-025-65978-4

Figure Lengend Snippet: a Relative mRNA levels of Fgfr1-4 in total renal tissue lysates from C57BL/6 mice ( n = 16). b , c Immunofluorescence images depicting FGFR1 distribution (green) in normal renal tissues from ( b ) C57BL/6 mice and ( c ) human samples. White dashed lines delineate glomerular boundaries. d , e High-magnification images showing co-localization of FGFR1 (green) with the podocyte marker podocin (red) in ( d ) mouse and ( e ) human glomeruli. f Schematic of podocyte-specific Fgfr1 knockout ( Fgfr1 -PKO), and Western blot of isolated glomeruli confirms loss of FGFR1 in Fgfr1 -PKO versus littermate controls. g , h Changes in ( g ) blood glucose levels and ( h ) BUN and UACR across treatment groups ( n = 6). i Renal excretion kinetics, GFR, and elimination half-life ( t 1/2 ) in mice from each group ( n = 3). j , k Histological examination and quantification of renal sections using H&E, PAS, and Masson’s trichrome staining ( n = 6). l , m immunohistochemistry and quantification of renal sections for the podocyte marker Nephrin from the indicated groups ( n = 6). n , o Representative images of isolated mouse glomeruli stained with WT-1 (green), TUNEL (green), and DHE (red), with quantification shown in ( o ) ( n = 6). Nuclei were counterstained with DAPI (blue). Data are presented as mean ± s.e.m. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 as determined by ordinary one-way ANOVA followed by Tukey’s multiple comparisons test ( a − o ); ns, not significant.

Article Snippet: The sections were then washed three times with PBS and incubated with Alexa Fluor 488- or 647-conjugated secondary antibodies (1:300, Cat. No. ab150073 or ab150115, Abcam) for 1 h. The sections were then incubated with DAPI (Southern Biotech, Birmingham) for 10 min. Co-staining for FGFR1 (1:1000, Cat. No. 60325, Proteintech) and NPHS2 (1:1000, Cat. No. 20384, Proteintech) was performed using a multiplex staining kit (Cat. No. AFIHC034, AiFang Biological) based on Tyramide Signal Amplification (TSA) technology.

Techniques: Immunofluorescence, Marker, Knock-Out, Western Blot, Isolation, Staining, Immunohistochemistry, TUNEL Assay

Journal: Nature Communications

Article Title: FGF4-FGFR1 signaling promotes podocyte survival and glomerular function to ameliorate diabetic kidney disease in male mice

doi: 10.1038/s41467-025-65978-4

Figure Lengend Snippet: a , b Human podocytes were isolated from the urine of DKD patients and exposed to high glucose conditions with or without rFGF4 treatment. a Representative images of TUNEL staining (green) and quantification of TUNEL-positive cells from the indicated groups ( n = 6). b Immunofluorescence analysis of tubulin (green) and quantification of the aspect ratio (width to height ratio) of human podocytes ( n = 6). White dashed lines mark the edge of the cells. c Representative images and quantification of isolated human glomeruli stained for WT-1 (green), TUNEL (green), DHE (red), and Nrf-2 (pink). Nuclei were counterstained with DAPI (blue). d Western blot and quantitative analysis of c-Cas3, CAT, SOD-2, and Nrf-2 protein levels in human glomeruli from the indicated groups ( n = 4). β-actin served as a loading control. e Western blot analysis of the activation of AMPK-FOXO1 pathway components in total glomerular tissue lysate. The ratios of phosphorylated proteins to total proteins are shown as scatter plots ( n = 4). f Schematic showing that FGF4 protects against DKD by activating the FGFR1-AMPK-FOXO1 signaling axis. Data are presented as mean ± s.e.m. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 as determined by unpaired two-tailed Student’s t -test ( a , b ) or ordinary one-way ANOVA followed by Tukey’s multiple comparisons test ( c − e ); ns, not significant. c-Cas3, cleaved-Caspase3; CAT, Catalase.

Article Snippet: The sections were then washed three times with PBS and incubated with Alexa Fluor 488- or 647-conjugated secondary antibodies (1:300, Cat. No. ab150073 or ab150115, Abcam) for 1 h. The sections were then incubated with DAPI (Southern Biotech, Birmingham) for 10 min. Co-staining for FGFR1 (1:1000, Cat. No. 60325, Proteintech) and NPHS2 (1:1000, Cat. No. 20384, Proteintech) was performed using a multiplex staining kit (Cat. No. AFIHC034, AiFang Biological) based on Tyramide Signal Amplification (TSA) technology.

Techniques: Isolation, TUNEL Assay, Staining, Immunofluorescence, Western Blot, Control, Activation Assay, Two Tailed Test