Additional file 2 of Cancer cell-derived exosomal circUSP7 induces CD8+ T cell dysfunction and anti-PD1 resistance by regulating the miR-934/SHP2 axis in NSCLC
Creators: name: Chen, Shi-Wei
name: Zhu, Shu-Qiang
name: Pei, Xu
name: Qiu, Bai-Quan
name: Xiong, Dian
name: Long, Xiang
name: Lin, Kun
name: Lu, Feng
name: Xu, Jian-Jun
name: Wu, Yong-Bing
Publisher: name: figshare
Year: 2021
Subjects: subject: Biophysics
subject: Biochemistry
subject: Physical Sciences not elsewhere classified
subject: Cell Biology
subject: Molecular Biology
subject: Pharmacology
subject: Biotechnology
subject: Immunology
subject: FOS: Clinical medicine
subject: Developmental Biology
subject: Cancer
subject: Hematology

description: Additional file 2: Supplementary Figure 1. SHP2 and miR-934 expression along with cirUSP7 in NSCLC tissues were measured using RT-qPCR analysis according to groups include lymph node metastasis, tumor size, and intravascular cancer embolus. Supplementary Figure 2. circUSP7 expression in several NSCLC cell lines was measured using RT-qPCR analysis. Supplementary Figure 3. The expression of circUSP7 in NSCLC cell lines and its exosomes. a circUSP7 expression in the A549 and NCI-H1299 cells was modified by transfection of shRNA to cause interference, and circUSP7 expression in the NCI-H460 and 95D cells were modified by cDNA transfection. b circUSP7 silencing is accompanied by decreased exosomal circUSP7 in A549 and NCI-H1299 cells, and circUSP7 overexpression is accompanied by increased exosomal circUSP7 in NCI-H460 and 95D cells. The data are presented as the mean ± SD. **P &lt; 0.01. Supplementary Figure 4. The secretion of IFN-γ, TNF-α, Perforin, and Granzyme-B expression in CD8 + T cells co-cultured with exosomes derived from NCI-H1299 cells was modified by transfection of shRNA and 95D cells was modified by cDNA transfection by ELISA. Supplementary Figure 5. The relationship between circUSP7 and miR-934 expression in CD8 + T cells. a circUSP7 or miR-934 expression in CD8+T cells was co-cultured with exosome derived from NCI-H1299 cells was modified by transfection of shRNA and 95D cells were modified by cDNA transfection. b circUSP7 expression in CD8+T cells was modified by miR-934 mimics or shRNA transfection, miR-934 expression in miR-934-overexpressing or miR-934-silenced CD8+T cells. The data are presented as the mean ± SD. **P &lt; 0.01. Supplementary Figure 6. miR-934 affects the function of circUSP7 in CD8+T cells. a The expression of circUSP7 and miR-934 in CD8+T cells co-cultured with exosomes derived from NCI-H460 and 95D cells was modified by transfection of shRNA, and modified with miR-934 expression. b The expression of circUSP7 and miR-934 in CD8+T cells co-cultured with exosomes derived from NCI-H1299 and A549 cells was modified by transfection of cDNA, and modified with miR-934 expression. c The secretion of IFN-γ, TNF-α, Perforin, and Granzyme-B expression in CD8 + T cells co-cultured with exosomes derived from NCI-H460 cells was modified by cDNA transfection with modified circUSP7 and miR-934 expression. d The secretion of IFN-γ, TNF-α, Perforin, and Granzyme-B expression in CD8+T cells co-cultured with exosomes derived from A549 cells was modified by transfection of shRNA with modified circUSP7 and miR-934 expression. Supplementary Figure 7. a A negative correlation between circUSP7 and miR-934 was observed in the NSCLC tissues (R2 = 0.1282; P &lt; 0.0001). b A negative correlation between SHP2 mRNA and miR-934 was observed in the NSCLC tissues (R2 = 0.2258; P &lt; 0.0001). c The PD1 antibody immunotherapy efficacy assessment using CT-based RECIST1.1. d and e The relationship between circUSP7 and miR-934, SHP2 and circUSP7, and SHP2 and miR-934 expression in patients with PD or PR + SD after anti-PD1 therapy. Supplementary Figure 8. The relationship between circUSP7 and Kras, p53 and ALK mutation status in NSCLC.