antimouse immunoglobulin g secondary antibodies  (Bio-Rad)

Journal: Kidney Research and Clinical Practice

Article Title: Tonsil-derived mesenchymal stem cells protect the kidney from gentamicin-induced acute kidney injury by incorporation into damaged renal tubules and amelioration of oxidative and endoplasmic reticulum stresses

doi: 10.23876/j.krcp.24.234

Figure Lengend Snippet: (A–D) GM (3 mM) induces apoptosis in cultured renal tubular cells (NRK-52E cells) assessed by FACScan analysis (A–C) and TdT-mediated dUTP nick-end labeling (TUNEL) staining (D: Control [a–c], GM [d–f], T-MSCs [g–i], GM + T-MSCs [j–l]). (F) Co-culture of T-MSCs (upper chamber) with NRK-52E (lower chamber) cells using transwell results in a significant decrease in apoptosis at 48 hours. GM-induced alteration of B-cell lymphoma 2 (Bcl-2)–associated X (Bax) and Bcl-2 is ameliorated by T-MSC co-culture. Representative early apoptotic (Annexin V [+]/PI [–]), late apoptotic (Annexin V [+]/PI [+]), and total apoptotic with quantitation bar (B, C). Representative TUNEL staining (D: green fluorescein with nuclear 4’,6-diamidino-2-phenylindole [DAPI] staining; magnification, ×200; scale bar = 100 μm) with quantitation bar (E) is shown. Representative Western blotting of Bax and Bcl-2 with quantitation bar is shown (F). Data are shown in box plots. FITC, fluorescein isothiocyanate; GM, gentamicin; PI, propidium iodide; T-MSC, tonsil-derived mesenchymal stem cell. * p < 0.05 vs. Control and T-MSCs, # p < 0.05 vs. GM without T-MSC.

Article Snippet: For immunofluorescence, sections were incubated with an anti-human nuclei antibody (1:200, MilliporeSigma) overnight, followed by an anti-mouse immunoglobulin G-fluorescein isothiocyanate (FITC) secondary antibody (1:200, Vector Laboratories), and mounted with DAPI (Vector Laboratories).

Techniques: Cell Culture, End Labeling, TUNEL Assay, Staining, Control, Co-Culture Assay, Quantitation Assay, Western Blot, Derivative Assay