TRF-2 Antibody

Knockdown Validated: TRF-2 Antibody [NB100-56694]

Knockdown Validated: TRF-2 Antibody [NB100-56694] - U2OS cells were transduced with a lentiviral vector coding for a Dox inducible control shRNA (shCtrl) or a shRNA against TRF-2 (shTRF2) and selected with puromycin +/- Dox for a week. One week post selection, +Dox cells were infected with HHV-6A for 48h and processed for IFA using anti-TRF-2 (green) and anti-IE2 (red). Cells with IE2 in punctate form and cells with large patchy IE2, likely to represent VRC, are shown. Nuclei are outlined by dashed lines. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/32320442/) licensed under a CC-BY license.

Western Blot: TRF-2 Antibody [NB100-56694] -

Western Blot: TRF-2 Antibody [NB100-56694] - TRF2 knockdown affect HHV-6A/B chromosomal integration.A) U2OS cells were transduced with lentiviral vectors expressing a scrambles shRNA (shCtrl) or a shTRF2. After a week of selection, cells were monitored for TRF2 expression by western blot. B) After a week of selection, shCtrl & shTRF2 treated cells were infected with HHV-6A or HHV-6B. After 30 days, DNA was isolated & the relative frequency of integration, relative to shCtrl set at 100%, estimated by ddPCR. *p<0.05. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32320442), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] -

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] - TRF2 expression in HHV-6A-infected U2OS cells.U2OS cells were infected with HHV-6A & analyzed for TRF2 & IE2 expression at 24h, 48h & 72h post-infection by dual color immunofluorescence. A) Representative immunofluorescence of TRF2 & IE2 expression in bystander & IE2 expressing cells at 24, 48h & 72h post infection. B) Mean relative TRF2 expression ± SD in uninfected (blue), IE2- (green-uninfected bystander) or IE2+ (red-infected) cells at 24h, 48h & 72h post infection. Each symbol represents the relative TRF2 expression from a single nucleus. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32320442), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] -

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] - Binding of TRF2 to viral DNA during HHV-6A/B infection.A) Schematic representation of the HHV-6A/B genome. The DR6 probe used for hybridization is shown in red. Uninfected & HHV-6A-infected HSB-2 cells (B-D) or uninfected & HHV-6B-infectd Molt-3 cells (E-F) were analyzed for TRF2 binding to viral DNA using ChIP. The input was hybridized with Alu probe to assess quantity of starting material. Anti-IgG (negative control), anti-PolII (positive control) or TRF2 antibodies were used for immunoprecipitation. B) QPCR detection of GAPDH DNA following ChIP. Results are expressed as fold increase over control IgG. C & E) Eluted DNA was hybridized with 32P-labeled Alu, telomeric (TTAGGG)3 or HHV-6A (DR6) probes. After hybridization the membranes were washed & exposed to X-ray films. D & F) Densitometric analysis of relative binding of TRF2 to telomeric & viral DNA. Results of one experiment representative of three are presented & are expressed as signal after normalization to input. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32320442), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] -

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] - TRF2 is required for IE2 localization with telomeres.U2OS cells were transduced with a lentiviral vector coding for a Dox inducible control shRNA (shCtrl) or a shRNA against TRF2 (shTRF2) & selected with puromycin +/- Dox for a week. A) Western blot analysis of TRF2 expression one week post selection. Membranes were also probed with anti-tubulin antibodies to show the input material loaded. B) One week post selection, +Dox cells were infected with HHV-6A for 48h & processed for IFA using anti-TRF2 (green) & anti-IE2 (red). Cells with IE2 in punctate form & cells with large patchy IE2, likely to represent VRC, are shown. Nuclei are outlined by dashed lines. C) The percentage of HHV-6A infected cells (from B) was estimated after counting a minimum of 700 cells & scoring the IE2+ ones. Results are expressed as mean %IE2+ cells ± SD. D) Mean percentage ± SD of IE2 localizing with telomeres in the presence (shCtrl +Dox) & absence (shTRF2 +Dox) of TRF2. Each dot represents the % of IE2 foci localizing with telomeres in one nucleus. ****p<0.0001. E) IF-FISH confocal images of shCtrl (+Dox) & shTRF2 (+Dox) cells analyzed for TRF2 (green), IE2 (red) & telomeres (cyan). Nuclei are outlined by dashed circles. Examples of IE2 localizing with telomeres (top row) or not found with telomeres (bottom row) are highlighted by the dashed polygons. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32320442), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] -

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] - Binding of TRF2 to viral DNA during HHV-6A/B infection.A) Schematic representation of the HHV-6A/B genome. The DR6 probe used for hybridization is shown in red. Uninfected & HHV-6A-infected HSB-2 cells (B-D) or uninfected & HHV-6B-infectd Molt-3 cells (E-F) were analyzed for TRF2 binding to viral DNA using ChIP. The input was hybridized with Alu probe to assess quantity of starting material. Anti-IgG (negative control), anti-PolII (positive control) or TRF2 antibodies were used for immunoprecipitation. B) QPCR detection of GAPDH DNA following ChIP. Results are expressed as fold increase over control IgG. C & E) Eluted DNA was hybridized with 32P-labeled Alu, telomeric (TTAGGG)3 or HHV-6A (DR6) probes. After hybridization the membranes were washed & exposed to X-ray films. D & F) Densitometric analysis of relative binding of TRF2 to telomeric & viral DNA. Results of one experiment representative of three are presented & are expressed as signal after normalization to input. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32320442), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] -

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] - A) A stick diagram of the IE2 protein with various domains identified is presented. B) Colocalization of HHV-6A IE2 protein with telomeres in the absence of viral DNA. U2OS cells were transfected with an empty vector, with IE2 expression vector or with IE2 delta1290–1500 expression vector. Forty-eight hours later cells were processed for dual color immunofluorescence. Telomeres were labeled in cyan & IE2 in red. Nuclei are outlined by dashed lines. Examples of IE2 colocalizing with telomeres are presented in a 3D view (white arrows). C) The graph represents the mean ± SD % of WT IE2 & delta1290–1500 IE2 localizing with telomeres. D) Lack of colocalization between HHV-6A p41 & telomeres in uninfected cells. U2OS cells were transfected with an empty vector or with a p41 expression vector. Forty-eight hours later cells were processed for dual color immunofluorescence. TRF2 was labeled green, p41 in red & nuclei outlined by a dashed line. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32320442), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] -

Immunocytochemistry/ Immunofluorescence: TRF-2 Antibody [NB100-56694] - Colocalization of shelterin complex proteins & HHV-6A IE2 protein at VRC & cellular telomeres.A) U2OS cells were mock-infected or infected with HHV-6A for 48h after which cells were processed for IF-FISH. Telomeres were labeled in magenta, TRF2 in green & IE2 in red. The panels in the middle row show images of cells with IE2 patches overlapping with large, diffuse TRF2 & telomeric staining (rectangles). The panels in the third row represent infected cells with punctate IE2 pattern colocalizing with TRF2 & telomeres (dashed squares). The colocalization of IE2, TRF2 & telomeres are shown in both 2D & 3D images. B) Uninfected & HHV-6A-infected U2OS cells were transfected with an empty vector, a myc-tagged-TRF1 expression vector. Forty-eight hours later cells were processed for IF-FISH. TRF1 was labeled in green & IE2 in red. Nuclei were stained with DAPI. Images on the far right show 2D colocalization of TRF1 with IE2. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/32320442), licensed under a CC-BY license. Not internally tested by Novus Biologicals.