Journal: Life Medicine

Article Title: HDAC8-selective inhibitor PCI-34051 protects against aortic dissection by attenuating ferroptosis of vascular smooth muscle cells

doi: 10.1093/lifemedi/lnag013

Figure Lengend Snippet: AP-1 (c-FOS and c-JUN) is a key regulator of PCI-34051 in the regulation of HASMC ferroptosis. (A) Volcano plot displaying the gene expression distribution and differentially expressed genes among the DMSO and PCI-34051 groups after treatment with CD. (B) Gene Ontology (GO) enrichment analysis showed that biological processes and cellular components were associated with these differentially expressed genes. (C, D) Western-blot analysis and quantification showing the protein levels of c-FOS and c-JUN in HASMCs after treatment with DMSO and PCI-34051 under the ferroptosis models induced by CD. β-Actin served as a loading control ( n = 4 per group). (E, F) Co-immunoprecipitation results showed that HDAC8 interacted with c-JUN in HASMCs. (G) Results of GST pull-down demonstrated a direct interaction between HDAC8 and c-JUN in HASMCs. Values are means ± SD; *** P < 0.001. BP, biological process. CC, cellular component. MF, molecular function.

Article Snippet: The antibodies applied in this study were: β-Actin (AC026, ABclonal), FSP1 (20886-1-AP, Proteintech), GPX4 (ab125066, Abcam), c-JUN (T55290F, ABmart), c-FOS (T56596F, ABmart), Flag (F1804, Sigma-Aldrich), HDAC8 (17548-1-AP, Proteintech), SLC7A11 (26864-1-AP, Proteintech), 4-HNE (MAB3249-SP, Bio-techne), CD86 (13395-1-AP, Proteintech), α-SMA (ab7817, Abcam), α-SMA (GTX100034, Genetex), GST (AE001, Abclonal).

Techniques: Gene Expression, Western Blot, Control, Immunoprecipitation

Journal: Life Medicine

Article Title: HDAC8-selective inhibitor PCI-34051 protects against aortic dissection by attenuating ferroptosis of vascular smooth muscle cells

doi: 10.1093/lifemedi/lnag013

Figure Lengend Snippet: AP-1 (c-FOS and c-JUN) eliminated the effects of PCI-34051 on HASMC ferroptosis. All HASMCs were infected with lenti-Flag and lenti-c-FOS + lenti-c-JUN, and then these HASMCs were used for subsequent experiments. (A, B) The protein level of AP-1 (c-JUN and c-FOS) was detected by Western blot in HASMCs infected with lenti-Flag and lenti-c-JUN + lenti-c-FOS ( n = 4 per group). (C, D) The CCK8 assay showing the relative viability of HASMCs treated with DMSO and PCI-34051 after CD (C) and IKE (D) stimulation for the indicated time ( n = 5 per group). (E, F) Flow cytometry with propidium iodide (PI) staining displaying the percentage of PI-positive cells of HASMCs after treatment as described above ( n = 4 per group). (G, H) The LDH assay indicating the relative cell damage rate of HASMCs treated with described above ( n = 5 per group). (I–L) The ratio of oxidized BODIPY-C11/non-oxidized BODIPY-C11 fluorescence revealing the level of ROS of HASMCs treated with described above ( n = 4 per group). (M–P). 4-HNE immunofluorescence staining and quantitative analysis exhibiting the content of 4-HNE in HASMCs after treatment as described above ( n = 4 per group). (Q–T) Western-blot analysis and quantification performed to assess the protein levels of GPX4, FSP1, and SLC7A11 in AP-1 overexpressed HASMCs after DMSO and PCI-34051 treatment along with CD or IKE induction. β-Actin served as a loading control ( n = 4 per group). Values are means ± SD; *** P < 0.001, ** P < 0.01, * P < 0.05.

Article Snippet: The antibodies applied in this study were: β-Actin (AC026, ABclonal), FSP1 (20886-1-AP, Proteintech), GPX4 (ab125066, Abcam), c-JUN (T55290F, ABmart), c-FOS (T56596F, ABmart), Flag (F1804, Sigma-Aldrich), HDAC8 (17548-1-AP, Proteintech), SLC7A11 (26864-1-AP, Proteintech), 4-HNE (MAB3249-SP, Bio-techne), CD86 (13395-1-AP, Proteintech), α-SMA (ab7817, Abcam), α-SMA (GTX100034, Genetex), GST (AE001, Abclonal).

Techniques: Infection, Western Blot, CCK-8 Assay, Flow Cytometry, Staining, Lactate Dehydrogenase Assay, Fluorescence, Immunofluorescence, Control

Journal: Life Medicine

Article Title: HDAC8-selective inhibitor PCI-34051 protects against aortic dissection by attenuating ferroptosis of vascular smooth muscle cells

doi: 10.1093/lifemedi/lnag013

Figure Lengend Snippet: The working model of PCI-34051 regulating HASMC ferroptosis and aortic dissection in mice. PCI-34051 inhibits the ferroptosis of HASMC by affecting the interaction between HDAC8 and AP-1 (c-FOS and c-JUN) and further suppresses the occurrence and development of BAPN-induced aortic dissection in mice. (This figure is created with Biorender).

Article Snippet: The antibodies applied in this study were: β-Actin (AC026, ABclonal), FSP1 (20886-1-AP, Proteintech), GPX4 (ab125066, Abcam), c-JUN (T55290F, ABmart), c-FOS (T56596F, ABmart), Flag (F1804, Sigma-Aldrich), HDAC8 (17548-1-AP, Proteintech), SLC7A11 (26864-1-AP, Proteintech), 4-HNE (MAB3249-SP, Bio-techne), CD86 (13395-1-AP, Proteintech), α-SMA (ab7817, Abcam), α-SMA (GTX100034, Genetex), GST (AE001, Abclonal).

Techniques: Dissection