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. 2022 Jan 18;132(2):e153357. doi: 10.1172/JCI153357

Figure 5. Pseudokinase Trib1-mediated degradation of Cebpa is induced and required for Adrb3 downregulation.

Figure 5

(A and B) RNASeq was conducted on 3T3L1 adipocytes treated with 17 ng/mL TNF-α or 10 μM CL-316243 for 3 hours with or without 10 μM ESI09 (n = 3 per group, 1 sample from 3 independent experiments). (C and D) 3T3L1 adipocytes were treated with 10 μM CL-316243 or 17 ng/mL TNF-α (n = 3 per group). (E) 3T3L1 adipocytes were treated with Trib1 siRNA, then challenged with 1 μM CL-316243 or 17 ng/mL TNF-α. (F) 3T3L1 adipocytes treated with 17 ng/mL TNF-α for up to 48 hours (representative of n = 3 replicates). (G) 3T3L1 adipocytes were treated with siRNA against Trib1, then challenged with 1 μM CL-316243 or 17 ng/mL TNF-α for 3 hours (n = 3 per group). (H and I) Cebpα and Trib1 overexpression in 3T3L1 adipocytes was achieved by lentivirus followed by treatment with 1 μM CL-316243 or 17 ng/mL TNF-α for (H) 3 hours (n = 6 per group, from 2 independent experiments) or (I) 18 hours (n = 3 per group repeated once with similar results). *Significance compared with control or GFP unless otherwise specified. t test and FDRs applied to correct for multiple comparisons(A and B); 1-way ANOVA with Dunnett’s (C–E) or Tukey’s post hoc test (H). Error bars represent SEM. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.