PMC:6333292 / 45493-47033 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/6333292","sourcedb":"PMC","sourceid":"6333292","source_url":"https://www.ncbi.nlm.nih.gov/pmc/6333292","text":"Extended data Figure 8 Aspects of PD-L1 regulation by CMTM6 and STUB1.\n(a) V5-tagged PD-L1 was introduced into parental, CMTM6-overexpressing and CMTM6-deficient A375 cells. Cell lysates were denatured and then subjected to immunoprecipitation with anti-V5 antibody immobilized on protein G-coated beads. Immunoprecipitates were then analyzed by immunoblotting with anti-V5 antibody as a control for the experiments shown in Fig. 4e. Results of the FACS-based genetic screens in CMTM6 expressing and CMTM6 deficient HAP1 cells as shown in Fig. 1a (b) and in Fig. 3a (c), with the position of STUB1 indicated. (d) Relative expression of PD-L1, PD-L2 and the indicated PD-L1 – PD-L2 chimeric proteins in CMTM6 KD A375 cells as compared to matched control. Chimeras were detected with an anti PD-L1 or an anti PD-L2 antibody. (e) Schematic overview of the chimeric proteins analyzed. (f,g) 293T human embryonic kidney cells were co-transfected with a vector encoding either PD-L1, PD-L2 or the indicated chimeric protein, together with a vector encoding CMTM6. Cell lysates were denatured and subjected to immunoprecipitation with anti-flag antibody immobilized on protein G-coated beads. Lysates and immunoprecipitates were then analyzed by immunoblotting with the indicated antibodies. Data are representative of three (a,d), one (f) or two (g) independent experiments. Error bars represent s.d. of triplicates. MFI, median fluorescence intensity; KO, knockout; OE, overexpression; TM, transmembrane; IC, intracellular; EC, extracellular.\nE","divisions":[{"label":"Title","span":{"begin":24,"end":71}}],"tracks":[]}