Fig 1: Snail induces CXCL1 and CXCL2 via the NF-?B pathway and possibly via the direct binding to their promoters. a Western blotting of nuclear Snail, phosphorylated p65 (phospho-p65), p65 and RelB from HM-1, OVCAR8 and OVCA433 cells. HDAC1 was used as control. b Reverse transcription polymerase chain reaction (RT-PCR) to analyze the expression of Cxcl1 and Cxcl2 in HM-1-control and HM-1-shSnail cells (left), and the expression of CXCL1 and CXCL2 in OVCAR8-control and OVCAR8-shSnail cells (right), treated with or without BAY11-7082 (NF-?B inhibitor) at 10 µM for 24 h; n = 4. c Schematic representation of CXCL1 and CXCL2 promoter organization, and the corresponding luciferase reporter constructs pGL-CXCL1 (1523 bp: -1523 to +110 bp, 984 bp: -984 to +110 bp, 300 bp: -300 to +110 bp) and pGL-CXCL2 (1606bp: -1606 to +104 bp, 942 bp: -942 to +104 bp, 457 bp: -457 to +104 bp). TSS transcriptional start site, E1 exon1, and Luc luciferase. The black bars indicate E-boxes (CANNTG), which are the binding sites of Snail. d Luciferase reporter assays to analyze the activity of the pGL-CXCL1 and pGL-CXCL2 promoter constructs in 293FT-control and 293FT-shSnail cells. Relative luciferase activities are shown; n = 5. *P < 0.05, **P < 0.01, and ***P < 0.001 (one-way ANOVA with Tukey’s multiple comparisons test in b; unpaired t-test in d). Averaged data are presented as the mean ± SEM
Fig 2: SAHA alleviates brain damage in SAH rats by suppressing HSP70/HDAC1/TDP-43.a HDAC1 mRNA levels in rats after different treatments were determined by RT–qPCR. b Protein levels of HDAC1, acetyl-K, HSP70, and TDP-43 in rats with SAH after different treatments was determined by western blot analysis. c Nerve damage in rats with SAH after different treatments was determined by the modified Garcia behavior score. d The movement of rats with SAH after different treatments was determined by the rotarod test. e Memory impairment in rats with SAH after different treatments was determined by the Morris water maze test. f The degeneration of cortical neurons in rats with SAH after different treatments was determined by FJC staining. g Apoptosis in neurons was determined by TUNEL and NeuN double staining. h APP expression in rats with SAH after different treatments was determined by IHC staining. i Axonal damage in the brain tissue of rats with SAH after different treatments was observed by TEM. *P < 0.05, n = 6. The experiment was repeated three times independently.
Fig 3: Effects of HDACi and HDAC1 on YY1 expressionQRT-PCR and western blot analysis of HDAC1 and YY1 expressions in SMMC-7721 cells treated with HDACis (8 µM SAHA and 800 nM TSA for 48 hours) (A–D) and HEPG2 cells with HDAC1 overexpression (E–F) were performed. *P < 0.05.
Fig 4: Cryo-electron microscopy of the MiDAC complex.a Schematic of the domain structures of MIDEAS, HDAC1 and DNTTIP1: components of the MiDAC complex. b SDS-PAGE of the gel-filtration purification of the MiDAC complex on a Superdex-S200 column. c Section of an electron micrograph of the MiDAC complex. Scale bar: 20 nm. d 2D class averages of the dimer complex from Relion3. e 2D class averages of the tetramer complex from Relion3.
Fig 5: cPRC1 creates interactions between Polycomb domains.a, A representative Western blot analysis (n = 3) of nuclear extracts from the TetR-fusion lines used for Capture-C analysis probed with anti-Flag antibody to detect expression of the fusion proteins. HDAC1 is used as a loading control. b, ChIP-qPCR analysis of binding of the different TetR-fusion lines to the TetO array. Data are presented as mean value (n = 2) ±SD. Data points for individual replicates are shown. c, ChIP-qPCR analysis of binding of the CKM-Mediator complex to the TetO array in the TetR-CDK8, TetR-PCGF2, and TetR-GFP lines.. Data are presented as mean value (n = 2 for TetR-CDK8 and n = 3 for TetR-PCGF2 and TetR-GFP) ± SD. Data points for individual replicates are shown. d, Boxplot analysis of Capture-C mean normalised read counts and interaction scores in the TetR-fusion lines, looking at interactions with Polycomb domains (PCGF2-bound). Number of interactions is shown. Boxes show interquartile range, center line represents median, whiskers extend by 1.5x IQR or the most extreme point (whichever is closer to the median), while notches extend by 1.58x IQR/sqrt(n), giving a roughly 95% confidence interval for comparing medians. e, Snapshots showing Capture-C read count signal from TetR-CDK8, TetR-PCGF2 and TetR-GFP lines at a control locus. CDK8 and PCGF2 (cPRC1) ChIPseq signal is given as a reference. The Fli1 promoter bait is shown as a triangle and interactions created with surrounding cPRC1-bound sites are represented with arrowheads. f, A representative Western blot analysis of nuclear extracts (n = 3) from WT and cPRC1 KO ESCs probed with the indicated antibodies. TBP is used as a loading control. g, Metaplot analysis of CDK8 enrichment at CDK8 peaks (n = 24275) and Polycomb domains (n = 2097) in WT and cPRC1 KO ESCs. h, Heatmaps showing CDK8 ChIPseq signal at CDK8 peaks (n = 24275) and Polycomb domains (n = 2097) in WT and cPRC1 KO ESCs, sorted by decreasing CDK8 or RING1B signal, respectively. i, Boxplot analysis of Capture-C interaction scores from WT and cPRC1 KO ESCs showing interactions between Polycomb gene promoters and other Polycomb-domains (left), or non-Polycomb gene promoters and active sites (H3K27ac, right). Number of promoters (P) and interactions (int) is shown. Boxes show interquartile range, center line represents median, whiskers extend by 1.5x IQR or the most extreme point (whichever is closer to the median), while notches extend by 1.58x IQR/sqrt(n), giving a roughly 95% confidence interval for comparing medians. Source data
Supplier Page from Abcam for Anti-HDAC1 antibody [EPR460(2)]