Fig. 5: Spermidine actives CYLD to prompt RIP1 deubiquitination. | Cell Death & Disease

Fig. 5: Spermidine actives CYLD to prompt RIP1 deubiquitination.

From: Spermidine activates RIP1 deubiquitination to inhibit TNF-α-induced NF-κB/p65 signaling pathway in osteoarthritis

Fig. 5: Spermidine actives CYLD to prompt RIP1 deubiquitination.The alternative text for this image may have been generated using AI.

a Immunoblot analysis of the expression of UBA1 (E1), UBE2N (E2), TRIM21 (E3), CYLD in H-FLS treated by TNF-α for 2 min and spermidine for 1 h. b Immunofluorescence analysis of CYLD expression on the cytoplasm in H-FLS; scale bar, 20 μm. c RIP1 Ubiquitination, RIP1, CYLD, IKKβ, phosphorylated IKKβ (p-IKKβ), phosphorylated p65 (p-p65) was assessed by immunoprecipitation and immunoblotting in H-FLS transfected with HA-Ub and RIP1 (WT) plasmid, shRNA-Scramble (shRNA-SC) and treated with TNF-α (10 ng/mL) and spermidine (9 μM). d RIP1 Ubiquitination, RIP1, CYLD, IKKβ, phosphorylated IKKβ (p-IKKβ), phosphorylated p65 (p-p65) was assessed by immunoprecipitation and immunoblotting in H-FLS transfected with HA-Ub and RIP1 (WT) plasmid, shRNA-CYLD1 and treated with TNF-α (10 ng/mL) and spermidine (9 μM). e–g Quantification of protein expression for (a), (c), and (d) analysis of immunoprecipitation and immunoblotting respectively. Data are shown as mean ± SD, n = 3, *p < 0.05, **p < 0.01, ***p < 0.001, scale bar, 20 µm.

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