Fig. 1: PDZD8 interacts with Protrudin and is enriched in ER subdomains. | Nature Communications

Fig. 1: PDZD8 interacts with Protrudin and is enriched in ER subdomains.

From: PDZD8 interacts with Protrudin and Rab7 at ER-late endosome membrane contact sites associated with mitochondria

Fig. 1

a Endogenous PDZD8 and Protrudin interact. Volcano plots of mass-spectrometry based results from an analysis of immunoprecipitates using anti-PDZD8 (left panel) and anti-Protrudin (right panel) antibodies from crosslinked human HCT116 cell lysates showing significant potential interactors (in black), identified using one-sided Student’s t test (FDR 0.05, S0 = 0.1). PDZD8 and Protrudin are labeled in blue. Volcano plots represent experiments performed in three biological replicates for each condition. Source data are provided as a Source Data file. b Analysis of the PDZD8-GFP and Protrudin-mCherry interaction by immunoprecipitation from cell extracts. Extracts from HEK293T cells expressing Protrudin-mCherry, both Protrudin-mCherry and PDZD8-GFP or with no vector were immunoprecipitated with anti-GFP-Trap beads and western blot analysis was performed using anti-GFP, anti-Actin, and anti-Protrudin antibodies for detection of PDZD8-GFP, Actin, and Protrudin-mCherry (designated Protrudin-mCh), respectively. Source data are provided as a Source Data file. c PDZD8-GFP localizes to ER and to ER subdomains in cells. Top panel: representative images of U2OS cells expressing PDZD8-GFP (z-stack projection). Pdzd8-GFP (in green) and mitochondria (in red) labeled with MitoTracker DeepRed. Scale bar: 10 μm. Middle panel: U2OS cell expressing PDZD8-GFP (in green) shown full Z-stack projection (left) and a single plane (right). Lower panel: plots of pixel intensity versus distance using ImageJ software corresponding to the lines marked 1 and 2 on the single plane image in the middle panel.

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