Klotho Regulates 14-3-3ζ Monomerization and Binding to the ASK1 Signaling Complex in Response to Oxidative Stress
Fig 5
Effect of 14-3-3ζ knockdown on Klotho signaling via ASK1 and p38 MAPK pathway.
(A) Plots of phosphorylation levels of p38 MAPK in 14-3-3ζ-depleted HEK 293 cells. Cells were pre-treated with either 200 pM secreted Klotho or 20 mM NAC for 40 min before the addition of 5 μM ROT. Level of the phosphorylated protein was measured by Western blot using a phospho-14-3-3ζ (Ser-58) antibody. The same membrane was stripped and re-probed with the total 14-3-3ζ antibody to normalize for protein load. (B) Levels of Trx bound to ASK1 in 14-3-3ζ-depleted HEK 293 cells treated with secreted Klotho, or NAC, or buffer control for 40 min as described in text. The Western blot profile is shown beneath.