Fig 2: KAT5-induced FTL acetylation suppresses ferroptosis. (A,B) STRING analysis revealed TFCP2 interacted acyltransferase (A) and methyltransferases (B) (confidence = 0.4). (C) Co-immunoprecipitation experiments performed in PC9 cells using IgG, anti-KAT5 and anti-TFCP2 antibodies, and further analysis of KAT5, TFCP2 and GAPDH expression by IB. (D–G) The enrichments of H4K12Ac and H4K16Ac at –2k, TFCP2 (FOXA1) motif or 2k regions of FTL (D,F) or FTH (E,G) promoter were calculated as the percentage of input chromosomal DNA via ChIP using the corresponding antibodies in WT (D,E) or KAT5–/– (F, G) PC9 cells with erastin (10 µM) treated for indicated times. IgG was used as the parallel control. (H) FTH expression were measured using IB in WT, YAP–/–, TFCP2–/– and KAT5–/– PC9 cells with erastin (10 µM) treatment for indicated hours. (I–K) Labile iron, cell death and lipid ROS were measured in PC9 cells with or without KAT5 overexpression combined with FTL knockdown before erastin (10 µM, 24 h or 16 h) treatment. (L,M) Labile iron, cell death and lipid ROS generation were measured in PC9 and H1299 cells with or without FTL knocked down. Pentamidine (30 µM) and TH1834 (100 µM) were used to pretreat cells for 8 h before further treated with erastin (10 µM, 24 h). (N) The enrichments of YAP, TFCP2 and FOXA1 at –2k, TFCP2 (FOXA1) motif or 2k regions of FTL promoter were calculated as the percentage of input chromosomal DNA via ChIP using the corresponding antibodies in PC9 cells with or without KAT5 overexpressed. (O) Enrichment of YAP at TFCP2 (FOXA1)-binding motif within the FTL promoter in WT or KAT5–/– PC9 cells with or without YAP, TFCP2 and FOXA1 overexpressed was measured by ChIP-qPCR. (P) Co-IP experiments were performed using anti-YAP in PC9 cells with KAT5 overexpression or knockdown with or without erastin treatment (10 µM, 24 h). The YAP level in each co-IP samples was adjusted to the same protein content. The indicated proteins in co-IP samples or WCL were measured by IB. The data are shown as the mean ± SD from three biological replicates (including IB). *P < 0.05, **P < 0.01 indicates statistical significance. Data in (I–M,O) were analyzed using a one-way ANOVA test. Data in (N) were analyzed using student’s t-test.

Fig 3: Gene expression analysis in both contralateral control limbs and denervated limbs in wild type (WT, N = 5) and Rxfp2-/- mice (KO, N = 4), 3 days after denervation. Data in tibialis anterior and soleus muscle specimens are reported for Atrogin-1, MuRF-1, MUSA1, SMART, FBOX31, and TRAF6 genes, whose expression is normalized on GAPDH as housekeeping. Data are reported as mean values ± standard error of the mean. Significance: P-values between compared conditions are reported. Significant differences (P < 0.05) are highlighted in bold.

Fig 4: TRIAP1 exhibits low expression in patients with GDM. The expression levels of TRIAP1 in peripheral blood samples obtained from 30 patients with GDM and subjects with healthy pregnancies were detected by (A) reverse transcription-quantitative PCR and (B) western blotting. (C) The ratio of TRIAP1/GAPDH was calculated and presented. All experiments were performed in triplicate. The results are presented as the mean ± standard deviation. **P<0.01 vs. control. GDM, gestational diabetes mellitus; TRIAP1, TP53-regulated inhibitor of apoptosis 1.

Fig 5: Apelin relived endothelial dysfunction dependent on NF?B pathway. (A) Representative images of western blot for ICAM-1, VCAM-1, VEGFR-2, E-cadherin, p-NF?B, NF?B and GAPDH after HG and apelin treatment in NCMECs. (B, C, D, E and F) Quantification of ICAM-1, VCAM-1, VEGFR-2, E-cadherin, and p-NF?B/NF?B in (A). Values represent the mean ± s.d. n = 3, *P < 0.05.