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Figure S1-S8 from Eradication of Hepatocellular Carcinoma by NKG2D-Based CAR-T Cells
Figure S1. NKG2DL levels in 50 pairs of normal tissues (N) and liver cancers (T). Data was obtained from The Cancer Genome Atlas (TCGA) database. Statistical differences were analyzed using paired Student's t-tests, *P {less than or equal to} 0.05, **P {less than or equal to} 0.01, ***P {less than or equal to} 0.001. Figure S2. Immunohistochemical analysis was performed to detect the expression of MICA and ULBP2 in Normal (N1-N4), Cirrhosis (C1-C7) and tumors (T) tissue, scale bar = 100 μm. Figure S3. T cells were isolated from peripheral blood using the RosetteSep{trade mark, serif} Human T Cell Enrichment Cocktail and the cell-surface expression of CD3 was analyzed by flow cytometry. The RAJI cell line was used as a negative control. Figure S4. SMMC7721 cells were cells infected with plko-shcoo2, plko-shMICA, and plko-shULBP2 lentivirus. Quantitative RT-PCR analysis showing the efficiency of MICA and ULBP2 knockdown in SMMC7721 cells. The shMICA-2# and shULBP2-2# was used to construct SMMC7721-shMICA and SMMC7721-shULBP2 cell lines, respectively. shCOO2 was used as negative control. The data are presented as the means {plus minus} SD. Figure S5. Growth curves of T cells isolated from healthy donor, patient 1 and patient 2. The data are presented as the means {plus minus} SD. P< 0.05 for the indicated cells. Figure S6. Flow cytometry analysis of the expression of CD19 and NKG2D CAR in transduced T cells isolated from healthy donor, patient 1 and patient 2. Figure S7. Proportions of CD4+ and CD8+ T cells in NTD, CD19-BBz CAR-T, and NKG2D-BBz CAR-T cells by flow cytometry. Figure S8. Proliferation (A) and apoptosis (B) of CD19-BBz CAR-T and NKG2D-BBz CAR-T cells at day 11 after transduction. The data are presented in as the means {plus minus} SD, ns, not significant.
