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. 2020 Jul 23;10(20):9332–9347. doi: 10.7150/thno.47137

Figure 5.

Figure 5

Targeting USP7 can activate the p38 MAPK pathway to reprogram TAMs. (A) The strategy for sorting TAMs in TME by flow cytometry. (B) Heat maps illustrating the differentially expressed M1- and M2-related genes in TAMs between the P5091 group and the Control group based on the results of RNA sequencing. (C) RT-PCR further verifying the differentially expressed genes of sorted TAMs in each group. Data are presented as the mean ± SEM (n = 3). (D) KEGG analysis identifying 20 most obviously enriched pathways based on the differentially expressed genes of the two groups. (E) Western blotting detection of the expression of JNK, p-JNK, ERK1/2, p-ERK1/2, p38, p-p38, and β-actin in IL-4/13-BMDM M2 cells treated with P5091 (10 µM) at the indicated time points. (F) Flow cytometry analysis of CFSE expression on the surface of CD8+T cells in the presence of conditioned medium of IL-4/13-induced BMDM M2 cells from various indicated treatments. Treatments indicated: DMSO stimulation, P5091 (10 µM) stimulation, P5091 (10 µM) stimulation in the presence of inhibitors of p38 (SB203580, 10 µM), JNK (SP600125, 10 µM), Erk1/2 (U0126-EtOH, 10 µM). Data are presented as the mean ± SEM (n = 4).