Journal: JCI Insight

Article Title: Epithelial innate immunity mediates tubular cell senescence after kidney injury

doi: 10.1172/jci.insight.125490

Figure Lengend Snippet: (A–C) SA-β-Gal staining of kidneys in 3 mouse models of AKI (FA, IRI, and CP) 28 days after injury, compared with controls and relative quantification (D–F). Scale bars: 500 μm. (G–I) Representative immunofluorescence confocal images of LAMNB1 28 days after FA, IRI, and CP, and relative digital image analysis qualification of LAMNB1-positive cells (J–L). Scale bars: 20 μm. Data are presented as mean ± SD. P values were calculated with 2-tailed Student’s t test. Ten images per mouse. The numbers of experimental mice are indicated in each panel.

Article Snippet: Frozen sections (8 μm) were air dried for 20 minutes and then incubated at 37°C for 12 to 16 hours in fresh SA-β-Gal staining solution with 1 mg/ml X-Gal (Teknova) in dimethylformamide, 40 mM citric acid/sodium phosphate (pH 6.0), 5 mM potassium ferrocyanide, 150 mM NaCl, and 2 mM MgCl 2 .

Techniques: Staining, Immunofluorescence

Journal: JCI Insight

Article Title: Epithelial innate immunity mediates tubular cell senescence after kidney injury

doi: 10.1172/jci.insight.125490

Figure Lengend Snippet: (A) Representative images of kidneys stained for SA-β-Gal activity at low (upper panels) and high (lower panels) magnification of Ksp-CreMyd88fl/fl mice compared with Ksp-CreMyd88+/+ controls 28 days after FA injury, and (B) corresponding quantification by digital image analysis. Scale bars: 500 μm (upper) and 200 μm (lower). (C) Representative immunofluorescence confocal images of Ksp-CreMyd88fl/fl mouse kidneys probed with an antibody against LAMNB1 compared with controls 28 days after FA injury, and (D) corresponding quantification. Scale bars: 20 μm. (E) Representative immunofluorescence confocal images of Ksp-CreMyd88fl/fl mouse kidneys probed with an antibody against the proliferative marker Ki67 compared with controls 28 days after FA injury, and (F) corresponding quantification. Scale bars: 20 μm. Numbers of experimental mice are reported in each panel. Ten images per mouse. Data are presented as mean ± SD. P values were calculated with 2-tailed Student’s t test.

Article Snippet: Frozen sections (8 μm) were air dried for 20 minutes and then incubated at 37°C for 12 to 16 hours in fresh SA-β-Gal staining solution with 1 mg/ml X-Gal (Teknova) in dimethylformamide, 40 mM citric acid/sodium phosphate (pH 6.0), 5 mM potassium ferrocyanide, 150 mM NaCl, and 2 mM MgCl 2 .

Techniques: Staining, Activity Assay, Immunofluorescence, Marker

Journal: JCI Insight

Article Title: Epithelial innate immunity mediates tubular cell senescence after kidney injury

doi: 10.1172/jci.insight.125490

Figure Lengend Snippet: (A) Schematic representation of the protocol for tamoxifen-induced expression of the Cre transgene 5 days after injury in KspCre-ERT2Myd88fl/fl mice. TMX, tamoxifen; PBS, phosphate-buffered saline. (B) Representative images of kidneys stained for SA-β-Gal activity at low (upper panels) and high (lower panels) magnification of KspCre-ERT2Myd88fl/fl mice compared with controls (KspCre-ERT2Myd88+/+) 28 days after FA injury, and (C) corresponding digital image analysis. Scale bars: 500 μm (upper) and 200 μm (lower). (D) Representative immunofluorescence confocal images of KspCre-ERT2Myd88fl/fl mouse kidneys probed with an antibody against LAMNB1 compared with controls 28 days after FA injury, and (E) corresponding quantification. Scale bars: 20 μm. (F) Representative immunofluorescence confocal images of KspCre-ERT2Myd88fl/fl mouse kidneys probed with an antibody against the proliferation marker Ki67 compared with controls 28 days after FA injury, and (G) corresponding quantification. Scale bars: 20 μm. Numbers of experimental mice are reported in each panel. Ten images per mouse. Data are presented as mean ± SD. P values were calculated with 2-tailed Student’s t test.

Article Snippet: Frozen sections (8 μm) were air dried for 20 minutes and then incubated at 37°C for 12 to 16 hours in fresh SA-β-Gal staining solution with 1 mg/ml X-Gal (Teknova) in dimethylformamide, 40 mM citric acid/sodium phosphate (pH 6.0), 5 mM potassium ferrocyanide, 150 mM NaCl, and 2 mM MgCl 2 .

Techniques: Expressing, Staining, Activity Assay, Immunofluorescence, Marker