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Figure 4 | Cell Communication and Signaling

Figure 4

From: Panx1 regulates neural stem and progenitor cell behaviours associated with cytoskeletal dynamics and interacts with multiple cytoskeletal elements

Figure 4

Panx1 interacts with the cytoskeletal proteins Arp3 and β-Actin. (A) Western blot of EGFP immunoprecipitations from Panx1EGFP or control EGFP overexpressing N2a cells. Panx1EGFP ran at the expected molecular weight of ~75 kDa (~50 kDa Panx1 + 25 kDa EGFP), and could be visualized with both Panx1 and GFP specific antibodies. Samples were analyzed by LC-MS/MS for Panx1EGFP specific interacting proteins. (B) Sequence coverage of the Panx1EGFP interactor, Arp3, from peptides identified in two of three independent replicates of N2a transfection, immunoprecipitation and LC-MS/MS. Peptides identified are highlighted in yellow, and cover 48.8% of the sequence. (C) Confocal micrograph of N2a cells overexpressing Panx1EGFP and immunostained for Arp3. Areas of overlap are indicated by arrowheads. Areas of high Panx1EGFP localized to cellular processes are indicated by arrows. (D) Confocal micrograph of N2a cells overexpressing Panx1EGFP and immunostained for the β-Actin. Areas of overlap are indicated by arrowheads. Areas of high Panx1EGFP localized to cellular processes are indicated by arrows. (E) Western blot of an immunoprecipitation for Panx1 from untransfected N2a cells. The expected Panx1 band is present at ~50 kDa, as well as a lower band that likely corresponds to another glycosylation species. Arp3 and β-Actin were confirmed to interact with endogenous Panx1.

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