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. 2020 Oct 23;3(12):e202000821. doi: 10.26508/lsa.202000821

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© 2020 Martins-Marques et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 3. — (A) Cx43 was immunoprecipitated (IP) from HEK293^Cx43+-derived EVs. Cx43 phosphorylation was evaluated with phospho-Ser, phospho-Thr, and phospho-Tyr antibodies. (B) Phosphorylation of Cx43-S373 and Cx43-S368 was evaluated in HEK293^Cx43+-derived EVs. (C) HEK293^Cx43+ cells were treated with PMA or vehicle for 30 min in EV-depleted medium. EV-Cx43 levels were evaluated by WB (n = 4). (D) Cx43 ubiquitination evaluated after IP of Cx43 from HEK293^Cx43+ cells. (E) HL-1 cells were treated with PMA or vehicle control for 30 min in EV-depleted medium. EV-Cx43 levels were assessed by WB (n = 4). (F) Cx43 ubiquitination evaluated after IP of Cx43 from HL-1 cells. (G) Cx43 ubiquitination evaluated after IP of Cx43 in HEK293^Cx43+-derived EVs. (H) WB analysis of EV-Cx43 derived from HEK293^Cx43+ cells knockdown for Tsg101 (siTsg101), incubated in EV-depleted medium for 8 h (n = 4). NP, non-phosphorylated Cx43; P2, phosphorylated Cx43.

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