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. 2022 Jun 21;119(26):e2202631119. doi: 10.1073/pnas.2202631119

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Copyright © 2022 the Author(s). Published by PNAS.

This open access article is distributed under Creative Commons Attribution-NonCommercial-NoDerivatives License 4.0 (CC BY-NC-ND).

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Fig. 4. — SENP1-regulated FGFR1 SUMOylation/deSUMOylation maintains the balance between FGF2/FGFR1 signaling and VEGFA/VEGFR2 signaling in ECs. (A–C) Representative blots showing FGF/FGFR1 signaling in samples in HMVECs with SENP1 knockdown (A), inactive SENP1-mutant overexpression (B), and SENP1-WT overexpression (C). Arrowhead indicates band of interest. (D and E) FGF2/FGFR1 signaling in HMVECs infected with Ad-FGFR1-WT or Ad-FGFR1-2KR after FGF2 stimulation at the indicated time points. Representative blots are shown in D with quantification in E. Arrowhead indicates band of interest. The normalized value of p-FGFR1/FGFR1 is presented as mean ± SEM from three independent experiments. *P ≤ 0.05; **P ≤ 0.01. (F and G) VEGFA/VEGFR2 signaling in HMVECs infected with Ad-FGFR1-WT or Ad-FGFR1-2KR after VEGFA stimulation at the indicated time points. Representative blots are shown in F with quantification in G. Arrowhead indicates band of interest. The normalized value of p-VEGFR2/VEGFR2 is presented as mean ± SEM from three independent experiments. **P ≤ 0.01. (H) Model for SENP1-regulated FGFR1 SUMOylation/deSUMOylation maintains the balance between FGF2/FGFR1 signaling and VEGFA/VEGFR2 signaling in ECs; the consequent mechanism following SENP1-regulated FGFR1 SUMOylation restrains additional association of FGFR1/FRS2α but facilitates VEGFR2/FRS2α complex formation in ECs under hypoxia (gray part, the same procedures as shown in Fig. 3I). : FRS2α; : PTPRG; : Phosphate; : SUMO1; : SENP1; : enhanced signaling; : restrained signaling. CTL, control.

Inline graphic — SENP1-regulated FGFR1 SUMOylation/deSUMOylation maintains the balance between FGF2/FGFR1 signaling and VEGFA/VEGFR2 signaling in ECs. (A–C) Representative blots showing FGF/FGFR1 signaling in samples in HMVECs with SENP1 knockdown (A), inactive SENP1-mutant overexpression (B), and SENP1-WT overexpression (C). Arrowhead indicates band of interest. (D and E) FGF2/FGFR1 signaling in HMVECs infected with Ad-FGFR1-WT or Ad-FGFR1-2KR after FGF2 stimulation at the indicated time points. Representative blots are shown in D with quantification in E. Arrowhead indicates band of interest. The normalized value of p-FGFR1/FGFR1 is presented as mean ± SEM from three independent experiments. *P ≤ 0.05; **P ≤ 0.01. (F and G) VEGFA/VEGFR2 signaling in HMVECs infected with Ad-FGFR1-WT or Ad-FGFR1-2KR after VEGFA stimulation at the indicated time points. Representative blots are shown in F with quantification in G. Arrowhead indicates band of interest. The normalized value of p-VEGFR2/VEGFR2 is presented as mean ± SEM from three independent experiments. **P ≤ 0.01. (H) Model for SENP1-regulated FGFR1 SUMOylation/deSUMOylation maintains the balance between FGF2/FGFR1 signaling and VEGFA/VEGFR2 signaling in ECs; the consequent mechanism following SENP1-regulated FGFR1 SUMOylation restrains additional association of FGFR1/FRS2α but facilitates VEGFR2/FRS2α complex formation in ECs under hypoxia (gray part, the same procedures as shown in Fig. 3I). : FRS2α; : PTPRG; : Phosphate; : SUMO1; : SENP1; : enhanced signaling; : restrained signaling. CTL, control.