Journal: Frontiers in Pharmacology

Article Title: Transcription factor MEF2D regulates aberrant expression of ACSL3 and enhances sorafenib resistance by inhibiting ferroptosis in HCC

doi: 10.3389/fphar.2024.1464852

Figure Lengend Snippet: The transcription factor MEF2D regulates aberrant expression of ACSL3 (A) GSEA revealed enrichment of the peroxisome pathway in MEF2D-silenced HCC cells. (B) mRNA expression levels of ACSL3 and other oxidative stress-related candidate genes were analyzed in lv-shMEF2D-infected HCC cells and compared with controls (C) In PLC/PRF/5 cells infected with Lv-MEF2D or Lv-GFP, the mRNA expression of ACSL3 was quantitatively detected by qPCR, and the protein expression level was detected by Western blotting. Vinculin protein was used as an internal control. (D) Bioinformatic analysis for putative MEF2D-binding sites in the regulatory regions of ACSL3 was shown (E) Luciferase expression driven by ACSL3 promoter regions was measured in PLC/PRF/5 and Huh7 cells infected with Lv-MEF2D or Lv-shMEF2D. The folding changes of luciferasee relative activity in Lv-MEF2D- and Lv-shMEF2D infected cells were normalized to Lv-GFP- and Lv-scramble- infected cells, respectively. The data represent the mean ± SDs of three independent experiments. (F) A ChIP assay was performed to detect the binding of MEF2D to the potential MRE identified in the promoter regions of ACSL3. The IgG-incubated and blank groups were considered as negative controls, whereas the input fraction was the positive control (G) The mRNA expression levels of MEF2D in HCC tissues and normal tissues were analyzed using data from the TCGA database (H) The correlation between ACSL3 and MEF2D expression in HCC was analyzed using data from the TCGA database. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: ACSL3 antibody (Santa cruz, 1:100, Cat# sc-166374), ACSL4 Rabbit pAb (ABclonal, 1:1,000, Cat#A6826), MEF2D antibody (Santa cruz, 1:500, Cat# sc-271153), GPX4 Monoclonal antibody (proteintech, 1:1,000, Cat No.: 67763-1-Ig), Ferritin heavy chain Polyclonal antibody (FTH1) (proteintech, 1:1,000, Cat No.: 11682-1-AP), Vinculin Monoclonal antibody (proteintech, 1:10,000, Cat No.: 66305-1-Ig).

Techniques: Expressing, Infection, Western Blot, Control, Binding Assay, Luciferase, Activity Assay, Incubation, Positive Control

Journal: Frontiers in Pharmacology

Article Title: Transcription factor MEF2D regulates aberrant expression of ACSL3 and enhances sorafenib resistance by inhibiting ferroptosis in HCC

doi: 10.3389/fphar.2024.1464852

Figure Lengend Snippet: The transcription factor MEF2D regulates aberrant expression of ACSL3 (A) GSEA revealed enrichment of the peroxisome pathway in MEF2D-silenced HCC cells. (B) mRNA expression levels of ACSL3 and other oxidative stress-related candidate genes were analyzed in lv-shMEF2D-infected HCC cells and compared with controls (C) In PLC/PRF/5 cells infected with Lv-MEF2D or Lv-GFP, the mRNA expression of ACSL3 was quantitatively detected by qPCR, and the protein expression level was detected by Western blotting. Vinculin protein was used as an internal control. (D) Bioinformatic analysis for putative MEF2D-binding sites in the regulatory regions of ACSL3 was shown (E) Luciferase expression driven by ACSL3 promoter regions was measured in PLC/PRF/5 and Huh7 cells infected with Lv-MEF2D or Lv-shMEF2D. The folding changes of luciferasee relative activity in Lv-MEF2D- and Lv-shMEF2D infected cells were normalized to Lv-GFP- and Lv-scramble- infected cells, respectively. The data represent the mean ± SDs of three independent experiments. (F) A ChIP assay was performed to detect the binding of MEF2D to the potential MRE identified in the promoter regions of ACSL3. The IgG-incubated and blank groups were considered as negative controls, whereas the input fraction was the positive control (G) The mRNA expression levels of MEF2D in HCC tissues and normal tissues were analyzed using data from the TCGA database (H) The correlation between ACSL3 and MEF2D expression in HCC was analyzed using data from the TCGA database. **** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05.

Article Snippet: Cell lysates were pre-clarified with protein A/G beads and then incubated overnight at 4°C with protein A/G beads coated with anti-MEF2D antibody (2 μg, Santa cruz).

Techniques: Expressing, Infection, Western Blot, Control, Binding Assay, Luciferase, Activity Assay, Incubation, Positive Control

Journal: Journal of Advanced Research

Article Title: Tetramethylpyrazine nitrone exerts neuroprotection via activation of PGC-1α/Nrf2 pathway in Parkinson’s disease models

doi: 10.1016/j.jare.2023.11.021

Figure Lengend Snippet: Information about primary antibodies used.

Article Snippet: MEF2D , Mouse monoclonal , IgG3 , WB, 1:500 IF, 1:200 , sc-271153 , Santa Cruz.

Techniques: