Fig. 2: OTUD1 plays a role in MAPK/JNK pathway regulation and physically interacts with ASK1. | Nature Communications

Fig. 2: OTUD1 plays a role in MAPK/JNK pathway regulation and physically interacts with ASK1.

From: Aggresome formation promotes ASK1/JNK signaling activation and stemness maintenance in ovarian cancer

Fig. 2

a Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the differentially expressed genes between sgRNA-OTUD1 and control cells (sgCtrl). The top 9 enriched pathways were listed. The bubble size indicates changed gene numbers and colors represent false discovery rate (P‐value). b The specific effect of OTUD1 on the phosphorylation levels of several key components of the MAPK signaling pathway (ERK/p-ERK, p38/p-p38, JNK/p-JNK) were determined. c The expression of CD24, FOS and several other reported JNK target genes (IL6 and ATF2), as well as a number of CSCs markers such as OCT4, NANOG and NOTCH1 in SKOV3 control or OTUD1 depleted cells, were analyzed by qPCR (n = 3). d A dox-inducible EGFP-OTUD1 expression system was established to evaluate the effects of OTUD1 on protein levels of JNK signal pathway key components (ASK1/p-ASK1, JNK/p-JNK). Dox doxorubicin. e Co-immunoprecipitation (Co-IP) was performed with Flag-OTUD1 WT and HA-ASK1 in 293T cells. f Co-immunoprecipitation (Co-IP) was performed with Flag-OTUD1 WT or Flag-OTUD1 C320A mutant and HA-ASK1 in 293T cells. C320A, OTUD1-C320A. g Schematic illustration of OTUD1 domain. OTU ovarian tumor protease, UIM ubiquitin-interacting motif, IDR intrinsically disordered protein region. h Co-immunoprecipitation (Co-IP) was performed with Flag-OTUD1 (aa 1–286 or aa 287–481 truncation) and HA-ASK1 in 293T cells. P values are calculated using one-way ANOVA (c). Representative of n = 3 independent experiments (b, d–f, h). Source data are provided as a Source Data file.

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