Proteomics and imaging-based analyses identify a putative clathrin light chain.

(A) Subcellular localization of the HA-epitope tagged GlCHC reporter in the cell cortex. (B) Exclusive spectral counts for all proteins detected at high stringency parameters in a native co-IP using the GlCHC-HA bait. (C) Co-labeling of HA-tagged Gl4259 and endogenous GlCHC shows considerable signal overlap in three dimensional reconstructions of image stacks (inset: co-localization scatter plot). (D-F) Single optical sections of GFP-tagged Gl4259-reporters (D) co-labeled with the fluid phase marker dextran-TxR (E) (F: merge image). (G, H) Surface rendering of optical sections from the cell shown in D-F, shows a more peripheral localization of Gl4259-GFP (green) compared to the signal for the PV-marker dextran-TxR (red) (G: ventral view, H: caudo-dorsal view). (I) Localization of the Gl4259-APEX2-2HA reporter to the PPI in TEM micrographs after 5min exposure to DAB. (J-M) FRAP analysis shows no recovery in the bleached ROI03 over time indicating no measurable turnover for the Gl4259-GFP reporter. Pre-bleach (K), post-bleach t0 (L), post-bleach t350 (M). RFU: relative fluorescence units.