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. Author manuscript; available in PMC: 2024 Dec 9.
Published in final edited form as: Cell Rep. 2019 Dec 10;29(11):3522–3538.e7. doi: 10.1016/j.celrep.2019.11.028

Figure 3. Altered Dynamics of the SENP7 Interactome in DSS Mice.

Figure 3

(A) Coomassie staining (left panel) and immunoblotting (right panel) of samples co-immunoprecipitated with anti-SENP7 or IgG from colonic lysates of control and DSS7 mice. Samples from (A) were in-gel digested and analyzed by ESI-tandem mass spectrometry (MS/MS) (n = 5 mice).

(B) Venn diagram displaying the number of individual proteins identified by MS/MS.

(C) The relative abundance of various signaling pathways based on MS/MS prepared using the Reactome database (shown as that of DSS-7 mice versus control). Gene Ontology is represented as fold enrichment. Probability was determined using a binomial statistic for the false discovery rate (FDR) and a p value cutoff of ≤ 0.05 significance level.

(D) Protein-protein interaction network for SENP7 and its interacting hub proteins based on the Boolean network model. The size of the node (circles) in the network represents the fold change identified by the LFQ algorithm, and the border of the nodes represents iBAQ-based fold change.

(E) Schematic of the steps involved in co-culture experiments using CT26 cells and total immune cells from MLNs.

(F) Population frequency γδ T cells was calculated and plotted (labeling: immune cells, cells from MLNs; immune cells + IL15, IL-15 treatment of cultured immune cells for 3 days; C1, immune cells co-cultured with CT26 cells transfected with empty FLAG vector; SENP7 WT O/E, SENP7 wild-type overexpressed CT26 in co-culture setup; SENP7 C992A O/E, catalytic mutant of SENP7; SENP7 WT O/E + anti-IL15, SENP7 overexpressed epithelial cells with IL-15 neutralization).

(G) ELISA for indicated cytokines from the supernatant of C1, SENP7 WT O/E, SENP7 WT O/E + Anti-IL15, and SENP C992A O/E cells described above. Each plot is representative of three independent experiments. Results are represented as mean+ SEM.

p values were determined by Student’s t test and one-way ANOVA with Tukey’s posttest (*p < 0.05; **p < 0.01; ***p < 0.001; ns, non-significant). See also Figure S2 and Tables S1, S2, and S3.