Journal: Journal of Biomedical Science

Article Title: Endothelin-1 induces connective tissue growth factor expression in human lung fibroblasts by disrupting HDAC2/Sin3A/MeCP2 corepressor complex

doi: 10.1186/s12929-023-00931-5

Figure Lengend Snippet: ET-1 induced a decrease in HDAC2 activity and acetylation of H3. A Cells were stimulated with ET-1 for 5, 10, or 30 min. The cell lysates were immunoprecipitated with antibody specific for HDAC2, and then HDAC activity was detected. Bars indicate values of the mean ± SEM ( n = 3). * p < 0.05 versus control group. B Cells were stimulated with ET-1 for 5, 10, 20, 30, or 60 min, and then the acetylation of histone H3 was evaluated using Western blotting. H3 was used as a loading control. Bars indicate mean ± SEM ( n = 4). * p < 0.05 versus control group. C Cells were transfected with either 0.5 or 1 μg of HDAC2-HA plasmid for 24 h and then treated with ET-1 for 20 min. Cells were subsequently lysed and immunoblotted with antibodies specific for histone H3, acetyl-H3, or HA. Bars indicate values of the mean ± SEM ( n = 4). * p < 0.05 versus ET-1-treated group. D Cells were transfected with 1 μg of HDAC7-GFP plasmid for 24 h and then treated with ET-1 for 20 min. Cells were subsequently lysed and immunoblotted with antibodies specific for histone H3, acetyl-H3, or GFP. Bars indicate values of the mean ± SEM ( n = 5)

Article Snippet: Antibodies specific for Sin3A, MeCP2, and GFP-tag, were purchased from Cell Signaling Technology (Danvers, MA, USA), and the anti-α-tubulin antibody was purchased from Transduction Laboratories (Lexington, KY, USA).

Techniques: Activity Assay, Immunoprecipitation, Control, Western Blot, Transfection, Plasmid Preparation

Journal: Oncotarget

Article Title: Modulation of estrogen related receptor alpha activity by the kinesin KIF17

doi: 10.18632/oncotarget.18104

Figure Lengend Snippet: A. Sequence of the KIF17-Tail. The 12 amino acid NR box peptide containing the LXXLL motif is underlined. B. Co-immunoprecipitation of GFP-ERR1 with myc-EV control, myc-KIF17-T or myc-KIF17T ΔNR expressed in HEK293 cells. Immunoprecipitates and total lysates were analyzed by immunoblot using anti-GFP and anti-myc IgG. C. , D. Luciferase reporter assays showing transcriptional activity of endogenous ERR1 in MCF7 cells expressing ERRE-Luc and myc-EV, myc-KIF17-T, myc-KIF17 ΔNR (panel C) or myc-KIF17 NR (panel D). E. Luciferase reporter assays showing transcriptional activity of endogenous ERR1 in parental LM2 and LM2 NR cells transfected with ERRE-Luc. F. Same as in panel E, but comparing cells transfected with control or ERR1 siRNAs. Graphs show normalized luminescence values pooled from ≥ 3 experiments performed in triplicate. Error = SEM. *** p < 0.05.

Article Snippet: Lysates were cleared using Protein-G sepharose beads (GE Healthcare) and incubated overnight with 6-8μg rabbit anti-GFP antibody (Novus Biologicals, NB 600-303) or mouse anti-myc antibody (Sigma M4439).

Techniques: Sequencing, Immunoprecipitation, Control, Western Blot, Luciferase, Activity Assay, Expressing, Transfection