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. 2021 Jan 14;11:1399. doi: 10.1038/s41598-021-80999-x

Figure 5.

Figure 5

SHP2 inhibition enhances MHC class I and PD-L1 expression in cancer cells through IFNγ signaling. (a) Flow cytometry analysis of cell surface staining of MHC class I in tumor cells after 6 days of co-culture of OVCAR-8 spheroids with PBMCs or tumor only. MHC class I-positive cells were gated. Percentage of MHC class I-positive population was labeled. (b) Flow cytometry analysis of cell surface staining of MHC class I in tumor spheroids treated with IFNγ of gradient concentration in the absence or presence of SHP099 and TNO155 for 6 days. MHC class I-positive cells were gated. Percentage of MHC class I-positive population was labeled. (c) MFI of MHC class I in tumor cells after 6 days co-culture of OVCAR-8-sgAAVS1 or OVCAR-8-sgIFNGR1 spheroids with PBMCs. Relative percentage of MFI of tumor MHC I (SHP099 treated over DMSO group) was labeled. (d) MFI fold change (stimulated/unstimulated) of MHC class I in tumor cells after 6 days co-culture of OVCAR-8-CAS9-sgPTPN11-1 or OVCAR-8-CAS9-sgPTPN11-2 spheroids with PBMCs (3 donors). OVCAR-8 cells were treated with or without doxycycline (100 ng/ml) for 5 days before co-culture. (e) Histogram of tumor cell surface PD-L1 level from 6 days of co-culture and tumor only. (f) Histogram of cell surface PD-L1 level in tumor spheroids treated with IFNγ of gradient concentration in the absence or presence of SHP099 (20 μM) and TNO155 (1 μM) for 6 days. (g) MFI of PD-L1 in tumor cells after 6 days co-culture of OVCAR-8-sgAAVS1 or OVCAR-8-sgIFNGR1 spheroids with PBMCs. Relative percentage of MFI of tumor PD-L1 (SHP099 treated over DMSO group) was labeled. (h) MFI of PD-L1 in tumor cells after 6 days co-culture of OVCAR-8-CAS9-sgPTPN11-1 or OVCAR-8-CAS9-sgPTPN11-2 spheroids with PBMCs (3 donors). OVCAR-8 cells were treated with or without doxycycline (100 ng/ml) for 5 days before co-culture. Flow cytometry data was analyzed and processed with FlowJo (Version 10.7.1, https://www.flowjo.com/solutions/flowjo/downloads/previous-versions).