Immunoblot analysis of whole-cell extracts and cell-surface proteins from TNFAIP3-/- NF-kB-DCK*NALM-6 cells after CRISPR-Cas9 gene knockouts with the indicated sgRNAs. Cell surface proteins were captured on streptavidin agarose after surface biotinylation. Where indicated, cells were treated with NGI-1 (1 or 10 M) or tunicamycin, 1 M for 24 h. Mobility shifts in proteins between the whole-cell fraction and the surface fraction may be due to different loading buffers used for these fractions (seeSTAR Methods).
