Exosome‐like nanoparticles from Mulberry bark prevent DSS‐induced colitis via the AhR/COPS8 pathway

A

Mass spectrometry (MS) analysis of aryl hydrocarbon receptor (AhR) pulldown protein from mulberry bark‐derived exosome‐like nanoparticles (MBELNs) and 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD)‐treated MC38 cell lysate. Data from three technical replicates are shown.

B

Co‐immunoprecipitation of COP9/COP9 Constitutive Photomorphogenic Homolog Subunit 8 (COPS8) bound AhR from MC38 cell lysate treated with MBELNs. Data are mean ± SEM from three biological replicates, **P < 0.01 using Student’s t‐test.

C

Western blot analysis showing expression of COPS8 in mice ileum crypts. Data are mean ± SEM of three biological replicates, **P < 0.01 using Student’s t‐test.

D

Representative images showing effect of MBELNs with and without TCDD (1 nm)/ carbobenzoxy‐Leu‐Leu‐leucinal (MG132; 20 µM) on co‐expression of AhR and COPS8 using immunofluorescence. Scale bar 10 μm, images are from three biological replicates.

E

Western blot showing MBELNs, TCDD, and MG132 effect on co‐expression of AhR and COPS8. The data represent as mean ± SEM from three biological replicates **P < 0.01, ***P < 0.001 using one‐way ANOVA.

F

Heat map showing MS analysis of AhR pulldown protein from MC38 cells treated with MBELNs in combination with proteasome inhibitor, MG132. Data are represented from three technical replicates.

G

Expression of Cullin 1 (CUL1) and neddylated CUL1 (N‐CUL1) following MBELN treatment using Western blot. Data are mean ± SEM from three biological replicates. **P < 0.01 using Student’s t‐test.

H, I

Western blot showing expression of COPS8 in wild‐type (Wild) and AhR‐KO MC38 cells and expression of AhR in wild and COPS8‐KO MC38 cells. MC38 cells were KO using CRISPR/ CAS9 KO plasmid transfection.

Figure 2. MBELN treatment leads to inducing the expression of COPS8 via the AhR signaling pathway.