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. Author manuscript; available in PMC: 2019 Nov 29.
Published in final edited form as: Cell. 2018 Nov 15;175(6):1492–1506.e19. doi: 10.1016/j.cell.2018.10.007

Figure 4. TIS11B is required for 3′UTR-mediated cell surface localization of CD47 and interaction of SET with CD47-LU.

Figure 4.

(A) Co-IP of endogenous SET using GFP-Trap after transfection of GFP-CD47-LU into HeLa cells stably expressing the indicated shRNAs. 2.5 % of input was loaded. TUBULIN was used as additional loading control. shRNA Ctrl, control shRNA.

(B) FACS analysis of endogenous CD47 in HeLa cells stably expressing the indicated shRNAs. Mean fluorescent intensity (MFI) values are shown in parentheses. Shown is a representative experiment. Right panel shows mean ± SD of five biological replicates. t-test, ***, P < 0.001.

(C) Sequence of TNFα ARE.

(D) RNA pulldown of indicated biotinylated RNA oligonucleotides and western blot analysis of endogenous proteins in HeLa cells. 2.5 % of input was loaded. Mock, no RNA was transfected. ARE-1 and ARE-2 differ in the restriction sites used for cloning. HNRNPA1 specifically binds to SU1. TUBULIN served as loading control.

(E) FACS analysis of GFP after transfection of the indicated constructs into HeLa cells, shown as in (B). Bottom panel shows surface and total GFP expression as mean ± SD of five biological replicates. t-test, ***, P < 0.001.

(F) Co-IP of endogenous SET using GFP-Trap after transfection of the indicated constructs into HeLa cells. 2.5 % of input was loaded.

(G) Western blot of the indicated endogenous proteins in HeLa cells stably expressing TIS11B shRNA1. See also Figure S3B–I.