Journal: BMC Nephrology

Article Title: Meprin β activity modulates cellular proliferation via trans-signaling IL-6-mediated AKT/ERK pathway in IR-induced kidney injury

doi: 10.1186/s12882-025-04224-x

Figure Lengend Snippet: PCNA protein expression in kidney tissue from wild type (WT) and meprin β deficient mice (βKO) at 96 h post-IR. The real-time PCR data showed that mRNA expression levels of PCNA increased in both genotypes subjected to IR ( A ). Values for PCNA mRNA levels were presented as fold change relative to control WT kidneys and normalized to GAPDH mRNA. Each value represents the mean ± SEM of triplicate combinations from 4 mice per group. Immunohistochemical staining for PCNA in kidney tubules in wild-type (WT) and meprin β knockout (βKO) kidneys ( B ). Immunostaining for PCNA in renal corpuscles in both genotypes ( C ). OD data were quantified using Image J analysis Software (ImageJ/Fiji 1.46) and analyzed by two-way ANOVA for 10 non-overlapping fields from renal tubular and renal corpuscle sections from each kidney. Images at 60 × magnification and the scale bar representing 20 µm. There was a significant increase in PCNA protein expression in select tubules in both genotypes, but only in βKO in renal corpuscle of kidneys subjected to IR. Immunofluorescence counterstaining of PCNA (red) in kidney tubules with the proximal tubule markers, meprin β (green) in wild-type (WT) and villin (green) in meprin β knockout (βKO) kidneys in renal tubules ( D ). DAPI was used to stain the nuclei (blue). Images at 60 × magnification with a scale bar representing 20 µm. PCNA was primarily expressed in the lumen of PTs in both genotype kidney sections at 96 h post-IR. Data is expressed as mean ± SEM with P values as indicated, P ≤ 0.05 are considered statistically significant

Article Snippet: The primary antibodies used for immunohistochemistry included Meprin β (goat polyclonal, R&D Systems, Cat# AF3300, 1:200), Villin (mouse monoclonal, Santa Cruz, Cat# sc-58,897, 1:200), KIM-1 (rabbit polyclonal, Abcam, Cat# ab47635, 1:100), IL-6 (rabbit monoclonal, Thermo Fisher, Cat# 416–7061-82, 1:400 with meprin β), PCNA (rabbit monoclonal, Cell Signaling, Cat# 13,110, 1:400), p-AKT (rabbit polyclonal, Cell Signaling, Cat# 9271, 1:100), and p-ERK1/2 (rabbit polyclonal, Cell Signaling, Cat# 4370, 1:200).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control, Immunohistochemical staining, Staining, Knock-Out, Immunostaining, Software, Immunofluorescence