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. 2020 Sep 14;130(10):5523–5535. doi: 10.1172/JCI140155

Figure 5. RIOK interacts with RARRES1 and TNF-α induces apoptosis through RARRES1 and RIOK1.

Figure 5

(A) Podocytes were cotransfected with myc-DDK–tagged RIOK1 (RIOK1-myc-DDK) with either control EGFP vector, WT RARRES1-V5, or cleavage mutant FFAF-V5. Lysates were immunoprecipitated with V5 antibody to pull down the RARRES1-interacting proteins and probed with FLAG and RIOK-1 antibodies. Input lysates were probed with FLAG, RIOK1, V5, and GAPDH antibodies. (B) Human podocytes were transfected with overexpression plasmids of RARRES1-V5 and/or RIOK1-myc-DDK. Western blot analysis was performed 24 hours after transfection for phospho-p53 (ser46), cleaved caspase-3, and RARRES1. (C) Podocytes stably transduced with lentiviral vector expressing scrambled control shRNA or RARRES1 shRNA were treated with TNF-α (10 ng/mL for 24 hours). Lysates were probed for phospho-p53 (Ser46), RARRES1, and cleaved caspase-3. Representative blots of 3 independent experiments are shown.