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. Author manuscript; available in PMC: 2024 Mar 18.
Published in final edited form as: Sci Transl Med. 2024 Jan 10;16(729):eadd2029. doi: 10.1126/scitranslmed.add2029

Fig. 4. KMT2E-AS1 induces HIF-2α activation and prevents HIF-2α degradation in driving endothelial pathophenotypes.

Fig. 4.

(A and B) HIF-2α protein in hypoxic human PAECs after knockdown of KMT2E-AS1 (a) and overexpression of KMT2E-AS1 (B) (n = 3 or 4; *P < 0.05, **P < 0.01, ***P < 0.001, one-way (A) or two-way (B) ANOVA followed by Bonferroni’s post hoc analysis; data represent mean ± SEM. (C and D) ELOC RNA (c) and protein (D) expression in human PAECs after hypoxia and KMT2E-AS1 knockdown (Fig. 3B) (n = 3 to 5; *P < 0.05, **P < 0.01, one-way ANOVA followed by Bonferroni’s post hoc analysis; data represent mean ± SEM). (E) HIF-2α protein in human PAECs after KMT2E-AS1 knockdown and MG132 treatment (n = 3; **P < 0.01, one-way ANOVA followed by Bonferroni’s post hoc analysis; data represent mean ± SEM). (F) apoptotic caspase 3/7 activity in human PAECs after KMT2E-AS1 knockdown under hypoxia (left) and KMT2E-AS1 overexpression under normoxia (right) (n = 3; **P < 0.01, ****P < 0.0001, unpaired Student’s t test; data represent mean ± SEM). (G) BrdU proliferative potential after KMT2E-AS1 knockdown in hypoxia (left) and forced KMT2E-AS1 expression in normoxia (right) (n = 3 to 5; *P < 0.05, **P < 0.01, unpaired Student’s t test; data represent mean ± SEM). (H and I) Scratch wound healing assay (h) measures migration of human PAECs after KMT2E-AS1 knockdown under hypoxia (left, I) and KMT2E-AS1 overexpression in normoxia (right, I) (n = 6; ***P < 0.001, ****P < 0.0001, unpaired Student’s t test; data represent mean ± SEM). (J and K) human PASMC contraction in gel matrix (J) with conditioned media from PAECs after knockdown of KMT2E-AS1 under hypoxia (K, left graph) and after forced expression of KMT2E-AS1 under normoxia (K, right graph) (n = 6; ***P < 0.001, unpaired Student’s t test; data represent mean ± SEM). (L) endothelin-1 in conditioned media from human PAECs after KMT2E knockdown under hypoxia (left) and forced KMT2E expression in normoxia (right) by enzyme linked immunosorbent assay (n = 3; ***P < 0.001, unpaired Student’s t test; data represent mean ± SEM).