Extended Data Fig. 7: eIF5B is necessary for PD-L1 upregulation in UROD depleted human lung cancer cells and the KLN205 syngeneic mouse model.
From: eIF5B drives integrated stress response-dependent translation of PD-L1 in lung cancer
a, Flow cytometry analysis of cell-surface PD-L1 in independent human lung cancer cells. Bar graph represents Mean Fluorescence Intensity of cell-surface PD-L1 in PD-L1 Low (Calu-6, H2030) and PD-L1 High (H358, H441, Hop62) cell lines. b, Western blot analysis of PD-L1 and eIF5B in Calu-6 and H2030 cells expressing a control or UROD sgRNA transfected with control or EIF5B siRNA. c, Western blot analysis of PD-L1 and eIF5B in H441, H358 and Hop62 cells expressing a control or UROD sgRNA transfected with control or EIF5B siRNA. Experiments in b-c were performed two independent times with similar results. Data from a representative experiment are shown. d, Western blot analysis of PD-L1 in KLN205 cells expressing a Scrambled shRNA or two independent Urod shRNAs. e, Western blot analysis of PD-L1, UROD, and eIF5B in KLN205 cells expressing a control shRNA or Urod shRNA with and without Eif5b shRNA. Experiments in d and e were performed two independent times with similar results. Data from a representative experiment are shown. f, Quantification of tumor volumes of KLN205 cells shown in (e) transplanted to syngeneic DBA/2 mice (n = 8 mice for Scrambled shRNA, n = 5 for Eif5b shRNA, n = 9 for Urod shRNA and n = 8 for Urod + Eif5b shRNA). Linear mixed model was used to assess statistical significance, as described in Methods. p value (Scrambled vs Urod shRNA) = 0.0002, p value (Urod shRNA vs Urod + Eif5b shRNA) = 4.9351e-05.
