Extended Data Fig. 1: Selection and large-scale purification of monoclonal CCT5 C-term edited cell lines.
From: Snapshots of actin and tubulin folding inside the TRiC chaperonin
(a) Anti-FLAG western blot (top) and stain-free imaging (bottom) of wild type HEK293T, mixed pool, and five monoclonal cell lines. (b) Size exclusion profile of large-scale TRiC purification with pooled fractions indicated by pink shading. (c) SDS-PAGE of fractions from size exclusion chromatography showing double-banding patterns of TRiC subunits, and tubulin co-elution with TRiC. (d) Anti-CCT western blots (left) and Coomassie visualisation (right) of monoclonal 3.B5 line showing presence of TRiC subunits. Coomassie gel bands were excised, digested with trypsin and analyzed using LC-MS/MS to confirm presence of TRiC subunits. (e) Negative stain micrograph showing well-defined TRiC particles (data representative of n = 1). The Western Blot experiments in a,d were carried out at least two times. Uncropped images and unprocessed scans in a,c,e are available as source data.
