Journal: Nature Communications

Article Title: Transcription stress at telomeres leads to cytosolic DNA release and paracrine senescence

doi: 10.1038/s41467-024-48443-6

Figure Lengend Snippet: A Immunofluorescence of 53BP1 and γH2AX with in situ hybridization of telomeric DNA (TelC) in wt and Tcea1 –/– MEFs. Arrows denote TIFs on telomeres. The graph depicts the mean percentage of cells presenting ≥5 TIFs (n = 3). B pATM protein levels in wt and Tcea1 -/- MEFs whole-cell extracts (n = 3). The graph depicts the total ATM-normalized protein expression levels (n.e.l.). C TRF1 protein levels in whole-cell extracts from wt and Tcea1 –/– MEFs. Fibrillarin was used to normalize protein expression levels (n.e.l., n = 4). D Trf1 mRNA levels in wt and Tcea1 -/- MEFs (n = 3). E ChIP signals of TRF1 protein (shown as percentage of input after IgG normalization) on telomeres of wt and Tcea1 -/- MEFs (n = 5). F Immunofluorescence of TRF1 with in situ hybridization of telomeric DNA (TelC) in wt and Tcea1 –/– MEFs. The graph depicts the mean fluorescence intensity (MFI) of TelC in Tcea1 –/– MEFs and wt controls (n = 4). G OxiDIP signals of 8-oxoG (shown as percentage of input after IgM normalization) on telomeres, GC-rich and AT-rich regions of untreated wt, H 2 O 2 -treated wt and untreated Tcea1 –/– MEFs (n = 3). H Immunofluorescence against 8-oxoG with in situ hybridization of telomeric DNA (TelC) in wt and Tcea1 –/– MEFs. White arrows indicate 8-oxoG signal on telomeres (n = 4). The graph depicts the 8-oxoG mean fluorescence intensity (MFI) on telomeric DNA (TelC) in Tcea1 –/– and wt MEFs. Data analysis was performed using two-tailed Student’s t test. All data are presented as mean values ± SEM. Unless otherwise indicated, n = biologically independent experiments and scale bars are set at 5μm. Source data are provided as a Source Data file.

Article Snippet: Antibodies against 53BP1 (NB100-304, IF: 1:200), ATM (NB100-220, WB: 1:500) and goat anti-mouse IgM 550 (NB120-9167R, IF: 1:200) were from Novus Biologicals.

Techniques: Immunofluorescence, In Situ Hybridization, Expressing, Fluorescence, Two Tailed Test

Journal: Journal of Biological Chemistry

Article Title: Nature-inspired design and evolution of anti-amyloid antibodies

doi: 10.1074/jbc.ra118.004731

Figure Lengend Snippet: Figure 5. Western blot analysis of the conformational and sequence specificity of the AF1 antibody. Aβ, tau, and α-synuclein [disaggregated (D) and fibril (F); 0.85 µg] were separated via SDS-PAGE and transferred to nitrocellulose membranes. The AF1-Fc antibody was used at 10 nM (1% milk). Sequence-specific antibodies against Aβ (NAB228, 1:1000 dilution, 1% milk), tau (1E1/A6, 1:5000 dilution, 1% milk) and α-synuclein (5C2, 1:10000 dilution, 1% milk) were also used as controls. Blots were imaged at short (15 s; AF1-Fc, NAB228) and long (5 min; 1E1/A6, 5C2) exposure times. The differences in the intensities of the molecular weight markers between different blots are due to the use of different secondary antibodies.

Article Snippet: Antibody binding was performed by incubating the blocked membranes with 10 mL of 100 nM scFv, 10 nM scFv-Fc fusion, 1:1000 dilution of NAB228 (2 mg/mL stock concentration; A8354, Sigma-Aldrich), 1E1/A6 (stock concentration unknown; 05-804, EMD Millipore) or 1:10000 of 5C2 (1 mg/mL stock concentration; NBP1-04321, Novus Biologicals).

Techniques: Western Blot, Sequencing, SDS Page, Molecular Weight

Journal: Chemical Reviews

Article Title: Smart and Multifunctional Materials Based on Electroactive Poly(vinylidene fluoride): Recent Advances and Opportunities in Sensors, Actuators, Energy, Environmental, and Biomedical Applications

doi: 10.1021/acs.chemrev.3c00196

Figure Lengend Snippet: (a) Schematic representation of Vitrovac/PVDF magnetoelectric composites produced in the study (A–F). Representation of the lateral view of the three-layer composites (G,H). (b) magnetoelectric response obtained from laminates with bilayer composite (sample A), three-layer magnetostrictive–piezoelectric–magnetostrictive (MPM) (sample G) composite, and three-layer PMP (sample H) configurations. (c) Magnetoelectric response obtained as a function of the different aspect ratios. Reproduced with permission from ref . Copyright 2015 Elsevier.

Article Snippet: Commercial magnetostrictive materials typically used for this application are Vitrovac (Fe 39 Ni 39 Mo 4 Si B 12 ), Terfenol-D (Tb 0.3 Dy 0.7 Fe 1.9–2 ), and Metglas (Fe 81 B 13.5 Si 3.5 C 2 ), which are usually used with a piezoelectric polymer, most often PVDF and copolymers., The magnetoelectric effect is quantified by the magnetoelectric coefficient ( a ME ) described in eq : where V ME is the induced magnetoelectric voltage, t p is the thickness of the piezoelectric material, and H AC is the applied magnetic field.

Techniques: Produced

Journal: Chemical Reviews

Article Title: Smart and Multifunctional Materials Based on Electroactive Poly(vinylidene fluoride): Recent Advances and Opportunities in Sensors, Actuators, Energy, Environmental, and Biomedical Applications

doi: 10.1021/acs.chemrev.3c00196

Figure Lengend Snippet: Summary of Representative PVDF-Based Sensors, Materials and Applications Areas, Indicating Also the Reported FOM: Sensitivity ( s ), Detection Range ( dr ), Standard Deviation ( sd ), Sensor Output ( so ), and Detection Limit ( dl )

Article Snippet: Commercial magnetostrictive materials typically used for this application are Vitrovac (Fe 39 Ni 39 Mo 4 Si B 12 ), Terfenol-D (Tb 0.3 Dy 0.7 Fe 1.9–2 ), and Metglas (Fe 81 B 13.5 Si 3.5 C 2 ), which are usually used with a piezoelectric polymer, most often PVDF and copolymers., The magnetoelectric effect is quantified by the magnetoelectric coefficient ( a ME ) described in eq : where V ME is the induced magnetoelectric voltage, t p is the thickness of the piezoelectric material, and H AC is the applied magnetic field.

Techniques: Standard Deviation, Colorimetric Assay, Shear

Journal: PLoS ONE

Article Title: Vesicular Location and Transport of S100A8 and S100A9 Proteins in Monocytoid Cells

doi: 10.1371/journal.pone.0145217

Figure Lengend Snippet: (A) S100A9 in the biotinylated plasma membrane fraction of THP-1 cells: non-biotinylated membrane fraction (lane a) and biotinylated cell surface fraction (lane b). (B) S100A8 expression in biotinylated THP-1 cells (same sample fractions as in (A). (C) Expression of S100A9 (left) and S100A8 (right) in the non-biotinylated membrane fraction (lane a) and biotinylated cell surface fraction (lane b) of human monocytes. Antibodies used were; α-S100A9 (NOVUS clone 1C10) and α-S100A8 (BMA; T1030; clone 8-5C2). 4 ng recombinant hS100A9 (rA9) was loaded.

Article Snippet: Then antibodies raised against human S100A8 (mAb 8-5C2 from BMA Biomedicals, Augst, Switzerland), S100A9 (mAb 43/8; produced at Active Biotech AB), S100A8/S100A9 (mAb 27E10; BMA Biomedicals, Augst, Switzerland), RAGE/Fc (MAB11451; R&D Systems Minneapolis, MN, USA) or TLR4 (MAB14781; R&D Systems, Minneapolis, MN, USA) were injected for 2–3 min in 10 mM Hepes, 0.15 M NaCl and 0.005% Surfactant P20 (HBS-P), pH 7.4, containing 1 mM Ca ++ and 10 μM Zn ++ .

Techniques: Clinical Proteomics, Membrane, Expressing, Recombinant

Journal: PLoS ONE

Article Title: Vesicular Location and Transport of S100A8 and S100A9 Proteins in Monocytoid Cells

doi: 10.1371/journal.pone.0145217

Figure Lengend Snippet: (A) Binding of anti-S100A8 (8-5C2), S100A9 (43/8) and S100A8/S100A9 (27E10) to SA captured biotinylated plasma membranes. Antibodies were injected at 6.25–50 nM for 2 min at 30 μL/min in assay buffer (HBS-P with 1 mM Ca ++ + 20 μM Zn ++ ). A 15 μL pulse of 10 mM glycine-HCl was used for regeneration. Capture level ~ 2.1 kRU. Kinetic evaluation was made using a 1:1 model for mAb 43/8 at 25 and 50 nM. An affinity of 7.0 nM ( k on 1.9 × 10 5 1/Ms; k off 1.3 × 10 −3 1/s) was calculated with maximal binding of 46 RU. (B) Injection of non-biotinylated THP-1 lysate over immobilized mAb 8-5C2, 43/8 and 27E10 (density ~ 3 kRU). Lysate, 0.45 mg/mL, diluted 5 to 40 times in assay buffer, injected for 3 min at 20 μL/min. Distinct binding was obtained when the THP-1 lysate was injected over the 27E10 surface whereas no binding was observed when lysate was passed over 8-5C2 and 43/8. (C) Binding of ten-fold diluted non-biotinylated THP-1 lysate to mAb 27E10 is inhibited after pre-incubation with 50 nM 27E10 but not with 8-5C2 or 43/8. Binding was expressed as % of response without antibody. (D) Sensorgrams obtained after injection of 50 nM mAb 8-5C2, 43/8 and 27E10 over SA captured (level ~ 1.1 kRU) solubilized membranes from surface biotinylated human monocytes. An affinity of ~ 6.1 nM (( k on 2.7 × 10 5 1/Ms; k off 1.6 × 10 −3 1/s) and maximal binding at ~ 12 RU was obtained after fit of sensorgram obtained with the 43/8 antibody to a 1:1 model. (E) Interaction of non-biotinylated lysate from human monocytes with 8-5C2, 43/8 and 27E10. Serially diluted lysate (5- to 40-fold in assay buffer) was injected over immobilized antibodies as in . Binding at high levels was demonstrated to mAb 27E10 (reaching steady state levels at ~ 1,000 RU) and was completely blocked when lysate (diluted 5-fold) was pre-incubated with 50 nM 27E10 but not with 8-5C2 or 43/8 (Fig 2C). Specific (i.e. displaceable by pre-incubation with anti-S100A9; data not shown) binding at much lower levels was also shown to 43/8 whereas non-specific binding was demonstrated to 8-5C2 (Fig 2E).

Article Snippet: Then antibodies raised against human S100A8 (mAb 8-5C2 from BMA Biomedicals, Augst, Switzerland), S100A9 (mAb 43/8; produced at Active Biotech AB), S100A8/S100A9 (mAb 27E10; BMA Biomedicals, Augst, Switzerland), RAGE/Fc (MAB11451; R&D Systems Minneapolis, MN, USA) or TLR4 (MAB14781; R&D Systems, Minneapolis, MN, USA) were injected for 2–3 min in 10 mM Hepes, 0.15 M NaCl and 0.005% Surfactant P20 (HBS-P), pH 7.4, containing 1 mM Ca ++ and 10 μM Zn ++ .

Techniques: Binding Assay, Clinical Proteomics, Injection, Incubation

Journal: PLoS ONE

Article Title: Vesicular Location and Transport of S100A8 and S100A9 Proteins in Monocytoid Cells

doi: 10.1371/journal.pone.0145217

Figure Lengend Snippet: THP1 cells were cultured for 48h in presence or absence of 10ng/ml TNFα and stained with antibodies against S100A9 (purple) (A). THP-1 cells were treated with either TNFα (B) or TNFα + IL10 (C); stained with anti-S100A8 (red), anti-S100A9 (purple) and anti-S100A8/S100A9 (green) antibodies. Arrows indicate the vesicles containing only hS100A9 or hS100A8. (D) Distribution of hS100A8 and hS100A9 proteins in THP-1 cells by Transmission Electron Microscopy. Black and white boxes represent magnification of areas where S100A8 and S100A9 hetero- or homodimers can be observed respectively. Further, black arrows highlight cytoplasmic vesicles or plasma membrane patches where hS100A9 (big dots) and hS100A8 (small dots) proteins co-localize, while red arrows point to vesicles containing only hS100A9 or hS100A8 homodimers.

Article Snippet: Then antibodies raised against human S100A8 (mAb 8-5C2 from BMA Biomedicals, Augst, Switzerland), S100A9 (mAb 43/8; produced at Active Biotech AB), S100A8/S100A9 (mAb 27E10; BMA Biomedicals, Augst, Switzerland), RAGE/Fc (MAB11451; R&D Systems Minneapolis, MN, USA) or TLR4 (MAB14781; R&D Systems, Minneapolis, MN, USA) were injected for 2–3 min in 10 mM Hepes, 0.15 M NaCl and 0.005% Surfactant P20 (HBS-P), pH 7.4, containing 1 mM Ca ++ and 10 μM Zn ++ .

Techniques: Cell Culture, Staining, Transmission Assay, Electron Microscopy, Clinical Proteomics, Membrane

Journal: PLoS ONE

Article Title: Vesicular Location and Transport of S100A8 and S100A9 Proteins in Monocytoid Cells

doi: 10.1371/journal.pone.0145217

Figure Lengend Snippet: THP-1 cells were cultured for 48h in presence or absence of 10 ng/ml TNFα. S100A8 and S100A9 mRNA expression was determined using qRT-PCR and was normalized to β-actin mRNA level. Data indicate mean ± SD of triplicate samples. Differences between treated and untreated groups are significant at ***P<0.001 by unpaired Student’s t test (A). Intracellular S100A8 and S100A9 protein expression was determined using Western blot (B). THP1 cells were treated with TNFα or left untreated. Cells were harvested either after 48 h (C) or after 24 h and 48 h (D); gently homogenized in homogenization buffer and post-nuclear supernatants (PNSs) were prepared. Using the PNSs as starting material, endosome enriched fractions were prepared using a discontinuous sucrose gradient centrifugation. Fractions were analysed by western blotting using the indicated antibodies. F1 is the interphase between 25% sucrose/homogenization buffer and F2 is the interphase between 35%/25% sucrose. (E) Immunofluorescence analysis of S100A9 localization in TNFα activated THP-1 cells; THP-1 cells cultured for 48 h with TNFα were fixed, permeabilized, stained with anti-S100A9 antibody (green) along with different vesicular marker antibodies anti-Rab5, anti-Rab7 and anti-cathepsin D (all of them marked in the red channel) and analyzed by confocal microscopy.

Article Snippet: Then antibodies raised against human S100A8 (mAb 8-5C2 from BMA Biomedicals, Augst, Switzerland), S100A9 (mAb 43/8; produced at Active Biotech AB), S100A8/S100A9 (mAb 27E10; BMA Biomedicals, Augst, Switzerland), RAGE/Fc (MAB11451; R&D Systems Minneapolis, MN, USA) or TLR4 (MAB14781; R&D Systems, Minneapolis, MN, USA) were injected for 2–3 min in 10 mM Hepes, 0.15 M NaCl and 0.005% Surfactant P20 (HBS-P), pH 7.4, containing 1 mM Ca ++ and 10 μM Zn ++ .

Techniques: Cell Culture, Expressing, Quantitative RT-PCR, Western Blot, Homogenization, Gradient Centrifugation, Immunofluorescence, Staining, Marker, Confocal Microscopy

Journal: PLoS ONE

Article Title: Vesicular Location and Transport of S100A8 and S100A9 Proteins in Monocytoid Cells

doi: 10.1371/journal.pone.0145217

Figure Lengend Snippet: THP-1 cells were cultured in triplicate either in the presence or absence of 10 ng/ml TNFα and TNFα + IL-10 (10 ng/ml). Culture supernatant was collected after 48h of incubation and S100A8/S100A9 dimer concentration was determined with ELISA following manufacturer’s protocol (A). Percent cell death was determined using LDH assay (B). In some experiments, THP-1 cells were stimulated with TNFα for 48 h in presence of methylamine (30 μM). In other experiments, cells were stimulated 12 h with TNFα without methylamine, followed by 36 h of culture with the drug. Alternatively, THP-1 cells were stimulated with TNFα; after 36 h, brefeldin A (5 μg/ml) was added and cells were cultured for another 12 h. Culture supernatant was collected after completion of incubation and S100A8/A9 heterodimer (C) and S100A9 (D) concentration was determined with ELISA following manufacturer’s protocol. Values are means of triplicates ± SD. Differences between various treatment groups are significant at **P<0.01, ***P<0.001 and ns = not significant by unpaired Student’s t test.

Article Snippet: Then antibodies raised against human S100A8 (mAb 8-5C2 from BMA Biomedicals, Augst, Switzerland), S100A9 (mAb 43/8; produced at Active Biotech AB), S100A8/S100A9 (mAb 27E10; BMA Biomedicals, Augst, Switzerland), RAGE/Fc (MAB11451; R&D Systems Minneapolis, MN, USA) or TLR4 (MAB14781; R&D Systems, Minneapolis, MN, USA) were injected for 2–3 min in 10 mM Hepes, 0.15 M NaCl and 0.005% Surfactant P20 (HBS-P), pH 7.4, containing 1 mM Ca ++ and 10 μM Zn ++ .

Techniques: Cell Culture, Incubation, Concentration Assay, Enzyme-linked Immunosorbent Assay, Lactate Dehydrogenase Assay

Journal: Cellular microbiology

Article Title: Chromatin-bound bacterial effector AnkA recruits HDAC1 and modifies host gene expression

doi: 10.1111/cmi.12461

Figure Lengend Snippet: (A) Schematic of AnkA truncated mutants encoded by the pCMV-tag2b and pT7-FLAG-MAT-Tag-2 vectors and predicted locations of protein domains. (B) Merged confocal immunofluorescence microscopy of AnkA truncations at 24 h post-transfection in HEK293T. (C) Merged confocal immunofluorescence microscopy at 24 h post-transfection in HEK293T with AnkA ΔN and AnkA ΔN2 truncations engineered with the SV40 large T-antigen NLS to re-establish nuclear localization.

Article Snippet: DNA was immunoprecipitated with the AnkA monoclonal antibody 5C2 (prepared against a recombinant AnkA devoid of most ANK repeats in the N-terminus), RNA polymerase II and HDAC1 antibodies (Active Motif, Carlsbad, CA).

Techniques: Immunofluorescence, Microscopy, Transfection

Journal: Cellular microbiology

Article Title: Chromatin-bound bacterial effector AnkA recruits HDAC1 and modifies host gene expression

doi: 10.1111/cmi.12461

Figure Lengend Snippet: (A) EMSA with rAnkA and rSATB1 incubated with the matrix attachment region (MAR) of IGH (wt = 5’-TCTTTAATTTCTAATATATTTAGAAttc-3’) known to bind SATB1, and with a mutated IGH MAR (mut = 5’-TCTTTAATTTCTACTGCTTTAGAAttc-3’). The underscored nucleotides indicate the areas where AT changes were made. Because AnkA is degraded during purification (Figure S1), several bands are present in EMSA. (B) MARs within the CYBB promoter were predicted with MARFinder MAR-Wiz 1.5 (blue line) using default values; a high likelihood was assigned with a value greater than the 0.6 cut-off threshold. AnkA binding along the entire CYBB promoter and downstream positions was assessed by chromatin immunoprecipitation-quantitative PCR (ChIP-qPCR) (red bars).

Article Snippet: DNA was immunoprecipitated with the AnkA monoclonal antibody 5C2 (prepared against a recombinant AnkA devoid of most ANK repeats in the N-terminus), RNA polymerase II and HDAC1 antibodies (Active Motif, Carlsbad, CA).

Techniques: Incubation, Purification, Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, ChIP-qPCR

Journal: Cellular microbiology

Article Title: Chromatin-bound bacterial effector AnkA recruits HDAC1 and modifies host gene expression

doi: 10.1111/cmi.12461

Figure Lengend Snippet: (A) Schematic of proximal CYBB promoter and regions where AnkA binds (shown in black boxes) (Garcia-Garcia et al., 2009b). (B) Double stranded wild-type and mutated probes used for EMSAs. The mutated bases are shown in red and are underlined. (C) EMSA with rAnkA and mutated CYBB probes. Arrows indicate probes with decreased AnkA binding. (D) Reporter β-galactosidase activity in uninfected HL-60 and A. phagocytophilum-infected HL-60 cells transfected with the indicated plasmids (n=3). *P<0.001 (Student’s t-test, α=0.05). Data are represented as mean ± SEM.

Article Snippet: DNA was immunoprecipitated with the AnkA monoclonal antibody 5C2 (prepared against a recombinant AnkA devoid of most ANK repeats in the N-terminus), RNA polymerase II and HDAC1 antibodies (Active Motif, Carlsbad, CA).

Techniques: Binding Assay, Activity Assay, Infection, Transfection

Journal: Cellular microbiology

Article Title: Chromatin-bound bacterial effector AnkA recruits HDAC1 and modifies host gene expression

doi: 10.1111/cmi.12461

Figure Lengend Snippet: (A) Wild-type CYBB probe A (from Figure 3B) EMSA assays with truncated AnkA mutants and labeled proximal CYBB promoter probe. (B) β-galactosidase reporter assay in HL-60 cells co-transfected with pCYBB-LacZ and plasmids expressing the truncated AnkA mutants (n=3); *P<0.05, **P<0.001 vs. FLAG control (Student’s t-test, two-sided α=0.05). Data are represented as mean ± s.d.

Article Snippet: DNA was immunoprecipitated with the AnkA monoclonal antibody 5C2 (prepared against a recombinant AnkA devoid of most ANK repeats in the N-terminus), RNA polymerase II and HDAC1 antibodies (Active Motif, Carlsbad, CA).

Techniques: Labeling, Reporter Assay, Transfection, Expressing, Control

Journal: Cellular microbiology

Article Title: Chromatin-bound bacterial effector AnkA recruits HDAC1 and modifies host gene expression

doi: 10.1111/cmi.12461

Figure Lengend Snippet: (A) DNA pull-down assay with uninfected (HL-60) or A. phagocytophilum-infected HL-60 cell (Aph) lysates and wild-type or mutated CYBB promoters. The first two lanes are controls that demonstrate the presence of AnkA and/or HDAC1 in samples before reaction with the CYBB or mutated probes. The remaining lanes show the results of interactions in the absence of AnkA (HL-60), with AnkA (Aph), with unlabeled promoter or mutated promoter probe (No Biotin CYBB or No Biotin mut-CYBB) that will bind the magnetic beads, and 5’ biotinylated promoter or mutated probe (5’ Biotin CYBB or Biotin mut-CYBB) that will not bind the magnetic beads. (B) Coomassie blue stain and (C) protein immunoblot analysis of eluted proteins from protein pull-down assay with HDAC1-FLAG overexpressed in HEK293T cells mixed with purified rAnkA. The overexpressed HDAC1 (60–65 kDA) readily bound to the FLAG mab matrix and was detected in proximity to large quantities of mouse immunoglobulin gamma heavy chains (Ighg; 55 kDa) dissociated from the matrix in the Laemmli buffer. HDAC1 detection increased simultaneously with a marked increase in AnkA binding compared to cells lacking HDAC1 overexpression. (D) Densitometry analysis of relative AnkA abundance in protein pull-down assay. (E) ChIP analysis of AnkA-, HDAC1- and RNA polymerase II-binding at the proximal CYBB promoter in AnkA-transfected HEK293T cells (n=3). Dashed line indicates 2-fold change in binding. (F) β-galactosidase reporter assay in HL-60 cells co-transfected with pCYBB-LacZ and pEYFP-N1 (black bars) or pEYFP-ankA (gray bars) and treated with 5 mM sodium butyrate HDAC inhibitor or 1µM mocetinostat HDAC1 inhibitor (n=3). Data normalized to empty vector, pEYFP-N1 control. *P<0.001 (Student’s t-test, α=0.05). Data are represented as mean ± SEM.

Article Snippet: DNA was immunoprecipitated with the AnkA monoclonal antibody 5C2 (prepared against a recombinant AnkA devoid of most ANK repeats in the N-terminus), RNA polymerase II and HDAC1 antibodies (Active Motif, Carlsbad, CA).

Techniques: Pull Down Assay, Infection, Magnetic Beads, Staining, Western Blot, Purification, Binding Assay, Over Expression, Transfection, Reporter Assay, Plasmid Preparation, Control

Journal: Cellular microbiology

Article Title: Chromatin-bound bacterial effector AnkA recruits HDAC1 and modifies host gene expression

doi: 10.1111/cmi.12461

Figure Lengend Snippet: The diagram depicts the specific regions in AnkA that mediate nuclear localization (red lines in first 4 ANK repeats show conserved hydrophobic domains at the 13th residues), regions most associated with DNA binding, specifically localized to the central ANK repeats deleted in AnkA-ΔN2 or to the AT hook-like C-terminal residues (RGR), and regions most strongly associated with promoter activity deleted in AnkA-ΔN2 and less in AnkA-ΔC.

Article Snippet: DNA was immunoprecipitated with the AnkA monoclonal antibody 5C2 (prepared against a recombinant AnkA devoid of most ANK repeats in the N-terminus), RNA polymerase II and HDAC1 antibodies (Active Motif, Carlsbad, CA).

Techniques: Binding Assay, Activity Assay