Figure 3. RIPK2 PROTAC reduces MDP-induced pro-inflammatory gene expression and iNOS protein levels in SIM-A9 cells.

SIM-A9 cells were pretreated with 1 μM RIPK2 PROTAC for 4 h followed by 20 h incubation with 100 ng/mL MDP. After that, cells were harvested for RNA and protein extraction. A) Graph shows RIPK2 degradation by the RIPK2 PROTAC completely reduced MDP-induced transcription of pro-inflammatory genes Nos2, Ptgs2, Il-1β, Tnfα, Il6, Ccl2, and Mmp9. B-E) Effects of RIPK2 PROTAC on MDP-induced iNOS, COX-2, and RIPK2 protein levels. qRT-PCR data are normalized to the reference genes Cyc1 and Rltr2aiap and represented as fold changes compared to MDP treatment, and immunoblot data are normalized to β-actin. One-way ANOVA with Bonferroni post-test, ***P<0.001. Data are represented as mean ± SEM from three independent experiments.