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. 2019 Sep 3;5:132. doi: 10.1038/s41420-019-0212-4

Fig. 1. PERP interacts with SERCA2b at ER–PM junctions.

Fig. 1

a Protein interacting partners of Halo-PERP were isolated from Mel202 cells using the HaloTag Mammalian Pull-Down System and identified by mass spectrometry. Maxquant intensity values for SERCA2 and PERP in three independent experiments are shown. b The interaction of PERP and SERCA2b was confirmed in two independent HaloTag pull-down experiments by immunoblotting as described in the Methods (proteins eluted from the resin by TEV enzymatic cleavage followed by a successive SDS-based elution). Figure shows two different exposures of SERCA2b panel to allow visualization of lower intensity bands. c PERP–SERCA2 complex formation was validated by SERCA2 IP in Mel202 cells expressing HaloTag or Halo-PERP using a HaloTag antibody. d Diagram of SERCA2a–c proteins, indicating the relative positions of the peptide identified by mass spectrometry (black) and the antibody immunogen sites (grey) used for validation of the SERCA2b–PERP interaction. Antibody 1 was used in HaloTag pull down validation (b) and antibody 2 was used for IP of SERCA2 (c). e Super-resolution images of HeLa BAC Venus-PERP cells co-expressing mCherry-SERCA2b, junctions between the ER and PM shown by arrows. Scale bar: 20 µm in full image and 5 µm in zoom panel