Research Paper Volume 15, Issue 14 pp 6658—6689

Figure 2. Inhibiting/Deleting NLRP3 in middle-aged mouse podocytes. (A–Y) Comparison of glomeruli from young and middle-aged mice treated with vehicle (Saline) or MCC950, as well as middle-aged Nlrp3-null mice by immunohistochemistry for NLRP3 (A–E), inflammasome downstream signaling components Caspase-1 (F–J), Interleukin-1β (IL-1β) (K–O), IL-18 (P–T) as well as the senescence marker p16 (U–Y). All immunostainings (brown color) were quantified using automated approaches, depicted as integral optical density/area (IOD/area) and analyzed by Student’s t-test statistics. (Z–DD) Staining for SA-β-Gal (blue) and nuclear red (red). SA-β-Gal is not detected in young mice (Z, DD), but increased in the glomerulus in saline treated middle aged mice, including podocytes (AA, DD). SA-β-Gal was lowered by MCC950 (BB, DD) and in age-matched NLRP3 null mice (CC, DD). Note that in all the conditions staining increased in glomeruli of middle-aged saline-treated and was lower in MCC950-treated (C, E) and in aged-matched NLRP3 null mice. The scale bars in the images correspond to 25 μm.