Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Testosterone Antagonizes Doxorubicin‐Induced Senescence of Cardiomyocytes

doi: 10.1161/JAHA.115.002383

Figure Lengend Snippet: T stimulates AKT phosphorylation and prevents the changes in TRF 2 and p53 levels caused by doxorubicin. A through C, Representative western blots (A) and band densitometry (B and C) for phosphorylated AKT in H9c2 cells preincubated with T or left untreated for 15 minutes followed by 20‐minute exposure to Dox or no treatment. D through F, Representative western blots (D) and band densitometry (E through G) of TRF 2 and p53 protein expression in H9c2 cells after no treatment or exposure to Dox with or without prior incubation with T, F and then T (F+T), or LY and then T ( LY +T). Data are shown as means+ SEM (3 observations per condition); P values are from Kruskal–Wallis test. Dunn's multiple comparisons test: ** P <0.01 vs no treatment. Ctr indicates control; Dox, doxorubicin; F, flutamide; LY , LY 294002 (phosphatidylinositol 3 kinase inhibitor); p, phosphorylated; T, testosterone; TRF2, telomere binding factor 2.

Article Snippet: The following primary antibodies were used (clones are indicated for monoclonal antibodies): anti–phosphorylated AKT (Ser 473 , clone D9E; Cell Signaling Technology), anti–phosphorylated AKT (Thr 308 ), anti‐AKT (H‐136), anti–phosphorylated NOS‐3 (Ser 1177 , 15E2; Santa Cruz Biotechnology), anti–NOS‐3 (6H2; Cell Signaling Technology), anti‐TRF2 (4S794.15; Imgenex, Novus Biologicals), anti–phosphorylated p53 (Ser 15 , D4S1H; Cell Signaling Technology), anti‐p53, anti‐GAPDH (FL‐335), anti‐AR (Millipore), and anti‐ERK1/2.

Techniques: Western Blot, Expressing, Incubation, Binding Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Testosterone Antagonizes Doxorubicin‐Induced Senescence of Cardiomyocytes

doi: 10.1161/JAHA.115.002383

Figure Lengend Snippet: NOS ‐3 is implicated in testosterone prevention of Dox‐triggered downregulation of TRF 2 and senescence. A, Band densitometry and representative western blot for phosphorylated NOS ‐3 in H9c2 cells pretreated or not with LY and then incubated with T for 15 or 15+20 minutes, as indicated. B, Representative western blot and band densitometry of TRF 2 protein expression in H9c2 cells after no treatment or exposure to Dox with or without prior incubation with T or LN and then T ( LN +T). C, Percentage of SA β‐galactosidase–positive H9c2 cells counted after the same treatments as in (B). Data are means+ SEM . Western blot experiments included 3 replicates for each condition, whereas SA β‐galactosidase was assessed 4 times for no treatment and Dox and 7 times for T or LN +T followed by Dox. Kruskal–Wallis P values are displayed above the graphs. Dunn's multiple comparisons test: **** P <0.001 vs no treatment. Ctr indicates control; Dox, doxorubicin; LN , l ‐NG‐nitroarginine methyl ester, or l ‐ NAME ; LY , LY 294002 (phosphatidylinositol 3 kinase inhibitor); NOS, nitric oxide synthase; p, phosphorylated; SA β‐gal, senescence‐associated β‐galactosidase; T, testosterone; TRF2, telomere binding factor 2.

Article Snippet: The following primary antibodies were used (clones are indicated for monoclonal antibodies): anti–phosphorylated AKT (Ser 473 , clone D9E; Cell Signaling Technology), anti–phosphorylated AKT (Thr 308 ), anti‐AKT (H‐136), anti–phosphorylated NOS‐3 (Ser 1177 , 15E2; Santa Cruz Biotechnology), anti–NOS‐3 (6H2; Cell Signaling Technology), anti‐TRF2 (4S794.15; Imgenex, Novus Biologicals), anti–phosphorylated p53 (Ser 15 , D4S1H; Cell Signaling Technology), anti‐p53, anti‐GAPDH (FL‐335), anti‐AR (Millipore), and anti‐ERK1/2.

Techniques: Western Blot, Incubation, Expressing, Binding Assay

Journal: Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease

Article Title: Testosterone Antagonizes Doxorubicin‐Induced Senescence of Cardiomyocytes

doi: 10.1161/JAHA.115.002383

Figure Lengend Snippet: T inhibits Dox‐caused senescence of primary neonatal cardiomyocytes via androgen receptor/phosphatidylinositol 3 kinase/nitric oxide synthase 3. A and B, Percentage of SA β‐gal–positive primary mouse neonatal cardiomyocytes after no treatment or exposure to Dox with or without prior incubation with T or E 2 (A) or T, F and then T (F+T), LY and then T ( LY +T), LN and then T ( LN +T), SB and then T ( SB +T), or SP and then T ( SP +T) (B). C, Representative pictures of the staining for SA β‐gal after the same treatments as in (A) and (B). Magnification is ×200, and bars correspond to 50 μm. D, Representative western blot and band densitometry of TRF 2 protein expression in primary mouse neonatal cardiomyocytes after no treatment or exposure to Dox with or without prior incubation with T, F and then T (F+T), or LY and then T ( LY +T). Means+ SEM and Kruskal–Wallis P values are shown in graphs (western blots: 3 observations per group; SA β‐gal: 7 assessments for no treatment, Dox, and T or E 2 followed by Dox; 4 for all the other conditions). Dunn's multiple comparisons test: * P <0.05, ** P <0.01, *** P <0.005, and **** P <0.001 vs no treatment; # P <0.05 vs Dox. Ctr indicates control; Dox, doxorubicin; E 2 , 17β‐estradiol; F, flutamide; LN , l ‐NG‐nitroarginine methyl ester, or l ‐ NAME (nitric oxide synthase inhibitor); LY , LY 294002 (phosphatidylinositol 3 kinase inhibitor); SA β‐gal, senescence‐associated β‐galactosidase; SB , SB 203580 (p38 inhibitor); SP , SP 600125 ( JNK inhibitor); T, testosterone; TRF2, telomere binding factor 2.

Article Snippet: The following primary antibodies were used (clones are indicated for monoclonal antibodies): anti–phosphorylated AKT (Ser 473 , clone D9E; Cell Signaling Technology), anti–phosphorylated AKT (Thr 308 ), anti‐AKT (H‐136), anti–phosphorylated NOS‐3 (Ser 1177 , 15E2; Santa Cruz Biotechnology), anti–NOS‐3 (6H2; Cell Signaling Technology), anti‐TRF2 (4S794.15; Imgenex, Novus Biologicals), anti–phosphorylated p53 (Ser 15 , D4S1H; Cell Signaling Technology), anti‐p53, anti‐GAPDH (FL‐335), anti‐AR (Millipore), and anti‐ERK1/2.

Techniques: Incubation, Staining, Western Blot, Expressing, Binding Assay

Journal: Molecular Biology of the Cell

Article Title: Telomeric DNA Mediates De Novo PML Body Formation

doi: 10.1091/mbc.E09-04-0309

Figure Lengend Snippet: Endogenous PML, Sp100 and Hausp accumulate at telomeric sites but not at centromeres in U2OS cells recovering from MMS treatment. (A) Immunofluorescence image of a U2OS cell treated with MMS, fixed and stained with anti-PML (green) and anti-TRF2 (red) antibodies. (B) Image of a U2OS cell that recovers from MMS treatment and is stained with antibodies against PML (green) and TRF2 (red). (C) Image of a HeLa cell that recovers from MMS treatment and is stained with antibodies against PML (green) and TRF2 (red). Arrows in B and C indicate the positions where PML colocalize or associate with TRF2 foci. (D) Localization of Sp100 at telomeric sites in a U2OS cell that recovers from MMS treatment. During recovery from MMS treatment, U2OS cells were fixed and stained with anti-Sp100 and anti-TRF2 antibodies. (E) Immunofluorescence image of a U2OS cell that recovers from MMS treatment. Sites where Hausp colocalize with telomeric DNA are indicated by arrows. (F) PML does not colocalize with centromeres in a U2OS cell that recovers from MMS treatment. After MMS treatment, U2OS cells were incubated in fresh medium, fixed and stained with anti-PML (green) and anti-CENPA (red) antibodies. All cell nuclei are counterstained with DAPI (blue).

Article Snippet: The following antibodies were used for immunofluorescence staining: mouse mAb 5E10 against PML (gift from R. van Driel, Amsterdam, The Netherlands), rabbit polyclonal antibody against PML (1130 directed against sequence: MEPAPARSPRPQQDP), rabbit polyclonal antibody against SP100 (ab1380, Chemicon, Temecula, CA), rabbit polyclonal antibody against Daxx (sc-7152, Santa Cruz Biotechnology, Santa Cruz, CA), rabbit polyclonal antibody against Hausp (A300–033A, Bethyl Laboratories, Montgomery, TX), mouse mAb against TRF1 (ab10579–50, Abcam, Cambridge, MA), mouse mAb against TRF2 (IMG-124, Imgenex, San Diego, CA), human autoimmune serum against centromeres (Antibodies Incorporated, Davis, CA), rabbit polyclonal antibody against γH2AX (A300–081A, Bethyl Laboratories), rabbit polyclonal antibody against 53BP1 (NB100–304, Novus Biologicals, Littleton, CO), and rabbit polyclonal antibody against SMC5 (A300–236A, Bethyl Laboratories).

Techniques: Immunofluorescence, Staining, Incubation

Journal: Molecular and Cellular Biology

Article Title: p53 Differentially Inhibits Cell Growth Depending on the Mechanism of Telomere Maintenance

doi: 10.1128/mcb.24.13.5967-5977.2004

Figure Lengend Snippet: FIG. 3. Expression of ts-p53TA perturbs APB frequency and causes an S phase delay in HIO107 ALT cells. (A) The frequency of APB-positive cells is increased two- to threefold when cell lines expressing either ts-p53WT or ts-p53TA are cultured at 32°C. APBs were detected by costaining the cells with a goat polyclonal antibody against PML and a rabbit polyclonal antibody (FC-08) against TRF2. (B) The HIO107-TA ALT cell line incorporates BrdU, but the cells accumulate in early S phase after 72 h at 32°C, while the HIO114 cell line is unaffected by growth at 32°C. The left panel shows a FACS analysis of BrdU intensity (y axis) versus DNA content (x axis). The right panel shows the cell cycle profiles generated from the data on the left of the figure (y axis, number of events; x axis, DNA content). (C) The parental HIO107 and the HIO107-TA cell lines were arrested by exposure to HU and, following release, cultured at either 32 or 39°C for the indicated time prior to harvest. The presence of active ts-p53TA prevents cells from progressing through S phase following the removal of HU. y axis, number of events; x axis, DNA content. (D) Quantitation of the cell cycle distribution of the populations shown in panel C. HIO107-TA cells accumulate in S phase for up to 24 h, suggesting that S phase is delayed when ts-p53TA is expressed. (E) Quantitation of the pattern of BrdU incorporation in HIO107-TA cells after 72 h of growth at 32°C. The majority of the cells are BrdU positive at 39°C, while at 32°C many of the cells are either BrdU negative or have punctate BrdU staining. (F) At 32°C, some HIO107-TA cells exhibit punctate BrdU staining which colocalizes with the telomeric protein TRF2 (arrow). The other two staining patterns also shown are diffuse nuclear staining or no BrdU incorporation.

Article Snippet: The precleared chromatin was incubated overnight at 4°C with the following primary antibodies: goat polyclonal human TRF2 (Imgenex), goat polyclonal p53 (C-19; Santa Cruz), or goat serum IgG (Santa Cruz).

Techniques: Expressing, Cell Culture, Generated, Quantitation Assay, BrdU Incorporation Assay, BrdU Staining, Staining

Journal: Molecular and Cellular Biology

Article Title: p53 Differentially Inhibits Cell Growth Depending on the Mechanism of Telomere Maintenance

doi: 10.1128/mcb.24.13.5967-5977.2004

Figure Lengend Snippet: FIG. 4. ChIP analysis. (A) Telomeric DNA is precipitated by anti- bodies against the telomeric binding proteins TRF2 but not by non- specific goat IgG, irrespective of culture conditions. Immunoprecipi- tation requires formaldehyde cross-linking of proteins to DNA. p53 is also associated with telomeric DNA in the HIO107-TA cell line but only at the permissive temperature of 32°C. Antibodies against TRF2 do not immunoprecipitate centromeric -satellite sequences. (B) Quantitation of the data shown in panel A. Percent input precip- itated (ppted) [(signal strength of telomeric DNA precipitated with the indicated specific antibody background signal obtained with nonspecific IgG)/input signal] 100. (C) Agarose gel of PCR products using primers to amplify the p53 binding site at position 1.4 kb within the p21WAF1 promoter. The template DNAs are identical to those interrogated for the experiment shown in panel C. The predicted product of 113 bp is only detected in DNA from the ChIP assay carried out with an antibody against p53 at 32°C. MW, PhiX HaeIII markers. Antibodies used for all panels are as follows: Gt TRF2, goat polyclonal antibody against TRF2 (Imgenex); Gt p53, goat polyclonal antibody against the carboxy terminus of human p53 (C-19; Santa Cruz); Gt IgG, goat IgG (Santa Cruz).

Article Snippet: The precleared chromatin was incubated overnight at 4°C with the following primary antibodies: goat polyclonal human TRF2 (Imgenex), goat polyclonal p53 (C-19; Santa Cruz), or goat serum IgG (Santa Cruz).

Techniques: Binding Assay, Quantitation Assay, Agarose Gel Electrophoresis

Journal: Oncotarget

Article Title: ERK1/2/MAPK pathway-dependent regulation of the telomeric factor TRF2

doi: 10.18632/oncotarget.10316

Figure Lengend Snippet: A. Equimolar amounts of GST, GST-ELK and His-TRF2 were incubated in the absence (−) or presence (+) of recombinant active ERK2. The phosphorylated proteins were detected after SDS PAGE using a specific anti-PX[phospho]SP antibody (pPXSP). Coomassie blue staining of the membrane is shown as a loading control. B. Alignment of TRF2 sequences from different mammalian species show the conservation of a MAPK phosphorylation consensus PXSP target site. The species and respective Genbank reference numbers corresponding to the sequences are reported. The conserved PXSP site is shown in bold and the conserved S residue underlined. C. Specificity of the immune serum using peptides containing phospho-S323. His-TRF2 was phosphorylated or not by recombinant active ERK2. The same amounts of proteins were submitted to immunoblotting analysis with the anti-pS323 antibody (pTRF2). Coomassie blue staining is shown as a loading control. D. A375 cells were stably transfected with WT-TRF2 or TRF2 S323A . Cells were treated (+) or not (−) with PD184352 (PD). Phosphorylated TRF2 was immunoprecipitated with the specific anti-pTRF2 antibody and detected by immunoblotting with an anti-TRF2 antibody (IP). Total TRF2 is shown as a loading control (input) and pERK1/2 as a control of PD184352 activity.

Article Snippet: Immuno-precipitation assays were performed on protein G sepharose beads with either 3 μg of the polyclonal phospho-TRF2 antibody or 2 μg of a TRF2 antibody (Imgenex IMG-124A) on 200 μg to 1 mg proteins, incubated for 16 hours at 4°C.

Techniques: Incubation, Recombinant, SDS Page, Staining, Membrane, Control, Phospho-proteomics, Residue, Western Blot, Stable Transfection, Transfection, Immunoprecipitation, Activity Assay

Journal: Oncotarget

Article Title: ERK1/2/MAPK pathway-dependent regulation of the telomeric factor TRF2

doi: 10.18632/oncotarget.10316

Figure Lengend Snippet: A. Immortalized BJ fibroblasts were serum deprived for 24 hours. BJ cells were then stimulated with 10% FCS for the indicated times. Phosphorylated forms of TRF2 were immunoprecipitated with the specific anti-pTRF2 antibodies and detected by immunoblotting with an anti-TRF2 antibody (IP p-TRF2). Total TRF2 and Hsp-90 are shown as loading controls and the phosphorylated forms of ERK1/2 as a control of serum-dependent activation of ERK1/2 (Input). B. Different tumor cells were tested for the presence of pTRF2 by immunoprecipitation in the presence (+) or absence (−) of PD184352 (PD) (Cal33, BJ-Ras (BJ-R), A375, U2OS). A short (Short exp.) and long (Long exp.) exposure of the blots are shown (IP p-TRF2). Total TRF2, Hsp90 and tubulin are shown as loading controls and the phosphorylated forms of ERK1/2 as a control of PD184352 activity (Input).

Article Snippet: Immuno-precipitation assays were performed on protein G sepharose beads with either 3 μg of the polyclonal phospho-TRF2 antibody or 2 μg of a TRF2 antibody (Imgenex IMG-124A) on 200 μg to 1 mg proteins, incubated for 16 hours at 4°C.

Techniques: Immunoprecipitation, Western Blot, Control, Activation Assay, Activity Assay

Journal: Oncotarget

Article Title: ERK1/2/MAPK pathway-dependent regulation of the telomeric factor TRF2

doi: 10.18632/oncotarget.10316

Figure Lengend Snippet: A. In situ proximity ligation assay (PLA) with anti-TRF2 and anti-pERK1/2 antibodies alone (negative controls, but in the presence of the two secondary antibodies) or in combination (TRF2 + pERK1/2) in A375 cells in the presence of DMSO or PD184352 (scale bars represent 10μm). A 3D image reconstruction after confocal microscopy imaging of PLA with a combination of anti-TRF2 and anti-pERK1/2 antibodies is also shown (scale bar represents 1μm). B. PLA with combined anti-TRF2 and anti-pERK antibodies in normal human skin tissue or cutaneous squamous cell carcinoma, normal lung or lung squamous cell carcinoma and normal cervix or cervical squamous cell carcinoma (scale bars represent 35μm). A 3D image reconstruction after confocal microscopy imaging of PLA with a combination of anti-TRF2 and anti-pERK1/2 antibodies in cutaneous squamous cell carcinoma is also shown (scale bar represents 1μm).

Article Snippet: Immuno-precipitation assays were performed on protein G sepharose beads with either 3 μg of the polyclonal phospho-TRF2 antibody or 2 μg of a TRF2 antibody (Imgenex IMG-124A) on 200 μg to 1 mg proteins, incubated for 16 hours at 4°C.

Techniques: In Situ, Proximity Ligation Assay, Confocal Microscopy, Imaging

Journal: Oncotarget

Article Title: ERK1/2/MAPK pathway-dependent regulation of the telomeric factor TRF2

doi: 10.18632/oncotarget.10316

Figure Lengend Snippet: A. A375, SKMel-51 and BJ-RAS cells were incubated in the presence of 50 μg/ml cycloheximide (CHX) for 16 hours in the presence or absence of PD184352 (PD). Total TRF2 and tubulin amounts were evaluated by immune-blotting. Tubulin and coomassie blue staining of the membrane (B) are shown as loading controls. B. Densitometric quantifications of the blots shown in A. The TRF2 expression level was normalized with three to four different loading controls and expressed relative to the control conditions (results are expressed as mean ± SD). One way ANOVA statistical analysis is included: * p<0.05; ** p<0.01; *** p<0.001). C. A375 cells over-expressing either TRF2-WT or TRF2-S323A were incubated in the presence of 50 μg/ml cycloheximide for 16 hours. Tubulin is shown as loading control. Densitometric quantification of the blot is shown (results are expressed as mean ± SD. One way ANOVA statistical analysis is included: * p<0.05; ** p<0.01; *** p<0.001).

Article Snippet: Immuno-precipitation assays were performed on protein G sepharose beads with either 3 μg of the polyclonal phospho-TRF2 antibody or 2 μg of a TRF2 antibody (Imgenex IMG-124A) on 200 μg to 1 mg proteins, incubated for 16 hours at 4°C.

Techniques: Incubation, Staining, Membrane, Expressing, Control

Journal: Oncotarget

Article Title: ERK1/2/MAPK pathway-dependent regulation of the telomeric factor TRF2

doi: 10.18632/oncotarget.10316

Figure Lengend Snippet: A. Confocal imaging of the co-staining of 53BP1 by immunofluorescence (red) and telomeres by fluorescent in situ hybridization (TelC-FITC, green) in A375 cells were the expression of WT-TRF2, TRF2 S323A or TRF2-ΔBΔM was induced by tetracycline (Tet) treatment. Colocalisation events were counted as Telomere dysfunction-Induced Foci (TIF, indicated with white arrows, scale bars represent 5μM) and the proportion of nuclei showing more than 3 TIF is indicated (right panel, results are expressed as mean ± SD, t-test statistical analysis is included: * p<0.05; ** p<0.01; *** p<0.001). B. The proportion of cells in each phase of the cell cycle was determined by DNA labeling with propidium iodide and FACS analysis. Sub G1 stands for cells with fragmented DNA, a hallmark of apoptosis. C. Conditional overexpression of different forms of TRF2 (WT, TRF2 S323A and TRF2 ΔBΔM ) was induced by tetracycline (Tet) in A375 cells. Seven days after tetracycline stimulation, cells were colored with giemsa blue. A close-up in TRF2 S323A overexpression well shows the few remaining cells at the end of the experiment. D. The cells were tested for b-galactosidase activity after seven days of tetracyclin induction (lower pictures). The percentage of β-galactosidase positive cells under tetracycline-induced conditions is specified below the images. E. Seven days after induction of the different forms of TRF2 by tetracycline (+), cells were tested for the presence of phosphorylated forms of p53. Actin and p53 are shown as loading controls.

Article Snippet: Immuno-precipitation assays were performed on protein G sepharose beads with either 3 μg of the polyclonal phospho-TRF2 antibody or 2 μg of a TRF2 antibody (Imgenex IMG-124A) on 200 μg to 1 mg proteins, incubated for 16 hours at 4°C.

Techniques: Imaging, Staining, Immunofluorescence, In Situ Hybridization, Expressing, DNA Labeling, Over Expression, Activity Assay

Journal: Oncotarget

Article Title: ERK1/2/MAPK pathway-dependent regulation of the telomeric factor TRF2

doi: 10.18632/oncotarget.10316

Figure Lengend Snippet: A. Control A375 cells or A375 cells conditionally expressing WT-TRF2 or TRF2 S323A were subcutaneously injected into nude mice. Doxycycline was added to the drinking water ten days after injection to induce the transgenes expression. The tumor volume is shown and results are expressed as mean ± SD (t-test statistical analysis is included: * p<0.05; ** p<0.01; *** p<0.001). B. Immunoblots showing the expression of TRF2 and actin (loading control) in tumor extracts prepared at the end of the tumor xenograft experiment.

Article Snippet: Immuno-precipitation assays were performed on protein G sepharose beads with either 3 μg of the polyclonal phospho-TRF2 antibody or 2 μg of a TRF2 antibody (Imgenex IMG-124A) on 200 μg to 1 mg proteins, incubated for 16 hours at 4°C.

Techniques: Control, Expressing, Injection, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Platinum Complexes Can Bind to Telomeres by Coordination

doi: 10.3390/ijms19071951

Figure Lengend Snippet: Quantification of TRF2 binding to telomeres by ChIP on A2780 cells treated with cisplatin, Pt-ttpy, or Pt-tpy for 8 h at 50 µM using anti-H3 and anti-TRF2 antibodies. The telomeric sequences immunoprecipitated by the anti-TRF2 or the anti-H3 antibodies were visualized after incubation of the dot blot membrane with a α 32 P radiolabelled 800 pb telomeric probe ( a ); signal normalization was performed after hybridization of the same membrane with α 32 P radiolabelled Alu sequences ( b ); two hundred nanograms of DNA were blotted for each ChIP sample. For the INPUT, 200, 100, and 50 µg of total DNA were blotted; ( c ) quantification of three ChIP experiments performed as in ( a ). Data were expressed as a percentage of the telomeric DNA signals in treated vs untreated cells. Quantitative values of the telomeric DNA signals are calculated as the ratio between the telomeric DNA signal precipitation and telomeric DNA signals in the INPUT for the same amount of blotted DNA. These values have been normalized to the amount of blotted DNA for each sample quantified by the non-specific Alu probe, following the formula: (telomere IP/telomere INPUT)/(Alu IP/Alu INPUT). (Means of at least three experiments) * Indicates a Mann and Withney test p -value p < 0.05 (GraphPad PRISM software, RITME, Paris, France).

Article Snippet: Immunoprecipitation was then performed with an anti-TRF2 polyclonal antibody (IMG-148A, IMGENEX).

Techniques: Binding Assay, Immunoprecipitation, Incubation, Dot Blot, Membrane, Hybridization, Software

Journal: International Journal of Molecular Sciences

Article Title: Platinum Complexes Can Bind to Telomeres by Coordination

doi: 10.3390/ijms19071951

Figure Lengend Snippet: TRF2 foci quantification detected by immunofluorescence on A2780 and A2780cis cells treated with cisplatin, Pt-ttpy, and Pt-tpy for 8 h at 50 µM; ( a ) A2780 cells were processed for immunofluorescence using antibodies against TRF2; ( b ) % of TRF2 foci after A2780 cell treatments with cisplatin, Pt-ttpy, and Pt-tpy ( c ) % of TRF2 foci after A2780cis cell treatments with cisplatin, Pt-ttpy, and Pt-tpy (mean of at least three experiments). * Indicates a Mann and Withney test p -value p < 0.05 (GraphPad PRISM software, RITME, Paris, France).

Article Snippet: Immunoprecipitation was then performed with an anti-TRF2 polyclonal antibody (IMG-148A, IMGENEX).

Techniques: Immunofluorescence, Software