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Proteintech rabbit polyclonal anti samhd1
Rabbit Polyclonal Anti Samhd1, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 90 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/rabbit+polyclonal+samhd1+antibody/SAMHD1+Antibody/pmc12184417__pnas%2E2424409122%2Esapp-226-27-30
Average 93 stars, based on 90 article reviews

rabbit polyclonal anti samhd1 - by Bioz Stars, 2026-09

93/100 stars

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Article Title: The impact of SAMHD1 expression and mutation status in mantle cell lymphoma: An analysis of the MCL Younger and Elderly trial.
Article Snippet: Immunohistochemical staining of SAMHD1 was performed on 3 μm thick sections of tissue microarrays (TMA) consisting of duplicate 0.6 mm cores using a semiautomated tissue stainer (LEICA) with conventional DAB staining. .. After heat-induced epitope retrieval at pH 9.0 the rabbit polyclonal SAMHD1 antibody (1:200; Proteintech) was incubated for 30 min, followed by washing and detection according to the manufacturer’s protocol. ..

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Journal: Cell reports

Article Title: Reprogramming of nucleotide metabolism by interferon confers dependence on the replication stress response pathway in pancreatic cancer cells

doi: 10.1016/j.celrep.2021.110236

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: anti-SAMHD1, rabbit polyclonal , Cell Signaling Technology , Cat#49158.

Techniques: Virus, Microarray, Recombinant, Lysis, Protease Inhibitor, Membrane, Saline, Autoradiography, Plasmid Preparation, Control, BIA-KA, Reverse Transcription, Enzyme-linked Immunosorbent Assay, Expressing, Software, Mass Spectrometry, Targeted Proteomics, Real-time Polymerase Chain Reaction, Live Cell Imaging

Figure 2. The impact of SAMHD1 expression on the failure-free survival of

Journal: International journal of cancer

Article Title: The impact of SAMHD1 expression and mutation status in mantle cell lymphoma: An analysis of the MCL Younger and Elderly trial.

doi: 10.1002/ijc.33202

Figure Lengend Snippet: Figure 2. The impact of SAMHD1 expression on the failure-free survival of

Article Snippet: After heat-induced epitope retrieval at pH 9.0 the rabbit polyclonal SAMHD1 antibody (1:200; Proteintech) was incubated for 30 min, followed by washing and detection according to the manufacturer’s protocol.

Techniques: Expressing

Primary human foreskin fibroblasts (HFFs) (panel a) or PMA-differentiated THP-1 cells (panel b) were infected with the indicated HCMV strains at an MOI of 1, or not infected (n.i.). At the indicated times after infection, real-time PCR was performed using primers specific for SAMHD1, or for the housekeeping gene GAPDH. Data from three (a) or two (b) independent experiments, expressed as fold change units ± SE, were normalized with GAPDH and referred to n.i. cells considered as calibrators and set ad 1. dpi, day post-infection; *, p < 0.05; **, p < 0.01.

Journal: PLoS Pathogens

Article Title: SAMHD1 phosphorylation and cytoplasmic relocalization after human cytomegalovirus infection limits its antiviral activity

doi: 10.1371/journal.ppat.1008855

Figure Lengend Snippet: Primary human foreskin fibroblasts (HFFs) (panel a) or PMA-differentiated THP-1 cells (panel b) were infected with the indicated HCMV strains at an MOI of 1, or not infected (n.i.). At the indicated times after infection, real-time PCR was performed using primers specific for SAMHD1, or for the housekeeping gene GAPDH. Data from three (a) or two (b) independent experiments, expressed as fold change units ± SE, were normalized with GAPDH and referred to n.i. cells considered as calibrators and set ad 1. dpi, day post-infection; *, p < 0.05; **, p < 0.01.

Article Snippet: The following antibodies were used in immunoblot: anti-SAMHD1 rabbit polyclonal antibody (Ab) (Proteintech); anti-SAMHD1 rabbit polyclonal Ab specific for the phosphorylated T592 residue (ProSci); rabbit polyclonal anti-p85 subunit of PI3K (N-SH2 domain) and mouse monoclonal antibody (mAb) anti-IE1/IE2 viral proteins (MAB810R; Merck Millipore); mAb anti-UL97 (kindly provided by Thomas Mertens, Ulm University Medical Center, Germany); rabbit anti-Cdk1 (#77055) and mAb anti-tubulin (both from Cell Signaling); rabbit anti-Cdk2 (sc-163) and mAb anti-lamin A (C-3) (both from Santa Cruz).

Techniques: Infection, Real-time Polymerase Chain Reaction

a) SAMHD1 levels were analyzed by immunoblotting in cell lysates of HFFs not infected (n.i.) or infected with AD169 at an MOI of 1 for 3 days. Expression of IE1/IE2 viral antigens was used as control for infection, while the p85 subunit of PI3K was used as loading control. A representative experiment out of six is shown. b) The relative amount of SAMHD1 protein, normalized to that of p85, was determined by densitometric analysis and is relative to that of n.i. cells, which was arbitrarily set as 1. Data are expressed as mean ± SE of eight independent experiments. c) SAMHD1 expression in HFFs not infected (n.i.) or infected with HCMV clinical isolates collected from infants affected by congenital HCMV infection (P1, P6, P7 and P10), or with Merlin. Data derive from one representative experiment out of two. d) SAMHD1 expression in HFFs, ARPE-19 and HMVEC cells infected with the indicated HCMV strains. Data derive from one representative experiment out of two, performed at the same time with all conditions. Numbers indicate the relative amount of SAMHD1 protein, determined as in panel b). e) The relative amount of SAMHD1 protein from HFFs infected with the indicated clinical strains was determined as in panel b). Data are expressed as mean ± SE of eight independent experiments. All data derive from cells infected at an MOI of 1 and harvested at 3 dpi. f) PMA-differentiated THP-1 cells not infected or infected with TR at an MOI of 1 for the indicated times. Expression of SAMHD1, viral antigens and p85 was evaluated as above. A representative experiment out of two is shown. g) SAMHD1 expression in HFFs induced to reach confluence (Con), serum-starved (Starv), proliferating (Prol), or infected with AD169 at an MOI of 1 for 3 days. Data derive from one representative experiment out of two. h) Antibody validation of the anti-SAMHD1 antibody by immunocapture MS. Proteins enriched by the antibody towards SAMHD1 ( y -axis) are plotted versus non-specific enrichment by one Rabbit IgG pool ( x- axis). Enriched proteins identified are quantified by label-free quantification and plotted as intensities (log 10 ). **, p < 0.01; ***, p < 0.001; ns, not significant.

Journal: PLoS Pathogens

Article Title: SAMHD1 phosphorylation and cytoplasmic relocalization after human cytomegalovirus infection limits its antiviral activity

doi: 10.1371/journal.ppat.1008855

Figure Lengend Snippet: a) SAMHD1 levels were analyzed by immunoblotting in cell lysates of HFFs not infected (n.i.) or infected with AD169 at an MOI of 1 for 3 days. Expression of IE1/IE2 viral antigens was used as control for infection, while the p85 subunit of PI3K was used as loading control. A representative experiment out of six is shown. b) The relative amount of SAMHD1 protein, normalized to that of p85, was determined by densitometric analysis and is relative to that of n.i. cells, which was arbitrarily set as 1. Data are expressed as mean ± SE of eight independent experiments. c) SAMHD1 expression in HFFs not infected (n.i.) or infected with HCMV clinical isolates collected from infants affected by congenital HCMV infection (P1, P6, P7 and P10), or with Merlin. Data derive from one representative experiment out of two. d) SAMHD1 expression in HFFs, ARPE-19 and HMVEC cells infected with the indicated HCMV strains. Data derive from one representative experiment out of two, performed at the same time with all conditions. Numbers indicate the relative amount of SAMHD1 protein, determined as in panel b). e) The relative amount of SAMHD1 protein from HFFs infected with the indicated clinical strains was determined as in panel b). Data are expressed as mean ± SE of eight independent experiments. All data derive from cells infected at an MOI of 1 and harvested at 3 dpi. f) PMA-differentiated THP-1 cells not infected or infected with TR at an MOI of 1 for the indicated times. Expression of SAMHD1, viral antigens and p85 was evaluated as above. A representative experiment out of two is shown. g) SAMHD1 expression in HFFs induced to reach confluence (Con), serum-starved (Starv), proliferating (Prol), or infected with AD169 at an MOI of 1 for 3 days. Data derive from one representative experiment out of two. h) Antibody validation of the anti-SAMHD1 antibody by immunocapture MS. Proteins enriched by the antibody towards SAMHD1 ( y -axis) are plotted versus non-specific enrichment by one Rabbit IgG pool ( x- axis). Enriched proteins identified are quantified by label-free quantification and plotted as intensities (log 10 ). **, p < 0.01; ***, p < 0.001; ns, not significant.

Techniques: Western Blot, Infection, Expressing, Control, Biomarker Discovery, Quantitative Proteomics

a) SAMHD1 phosphorylation at T592 (pT592) was analyzed by immunoblotting with a specific antibody in cell lysates of HFFs not infected or infected with AD169 at an MOI of 1 for the indicated time points. Expression of IE1/IE2 viral antigens was used as control for infection, while p85 was the loading control. A representative experiment out of six is shown. b) pT592-SAMHD1 was analyzed as in panel a), in parallel with an UVB-inactivated AD169 (UV), for the indicated time points. A representative experiment out of two is shown. c) pT592-SAMHD1 was analyzed as in panel a), in HFFs not infected or infected with the indicated strains (MOI 1, 3 dpi). A representative experiment out of three is shown. d) The relative amount of pT592-SAMHD1, normalized to that of p85, was determined by densitometric analysis and is relative to that of n.i. cells, which was arbitrarily set as 1. Data are expressed as mean ± SE of at least three independent experiments. *, p < 0.05. e) pT592-SAMHD1 levels in HFFs infected with HCMV clinical isolates (P1, P6, P7 and P10) or with Merlin. Data derive from one representative experiment out of two. *, unspecific band; black arrow, pT592-SAMHD1 specific band.

Journal: PLoS Pathogens

Article Title: SAMHD1 phosphorylation and cytoplasmic relocalization after human cytomegalovirus infection limits its antiviral activity

doi: 10.1371/journal.ppat.1008855

Figure Lengend Snippet: a) SAMHD1 phosphorylation at T592 (pT592) was analyzed by immunoblotting with a specific antibody in cell lysates of HFFs not infected or infected with AD169 at an MOI of 1 for the indicated time points. Expression of IE1/IE2 viral antigens was used as control for infection, while p85 was the loading control. A representative experiment out of six is shown. b) pT592-SAMHD1 was analyzed as in panel a), in parallel with an UVB-inactivated AD169 (UV), for the indicated time points. A representative experiment out of two is shown. c) pT592-SAMHD1 was analyzed as in panel a), in HFFs not infected or infected with the indicated strains (MOI 1, 3 dpi). A representative experiment out of three is shown. d) The relative amount of pT592-SAMHD1, normalized to that of p85, was determined by densitometric analysis and is relative to that of n.i. cells, which was arbitrarily set as 1. Data are expressed as mean ± SE of at least three independent experiments. *, p < 0.05. e) pT592-SAMHD1 levels in HFFs infected with HCMV clinical isolates (P1, P6, P7 and P10) or with Merlin. Data derive from one representative experiment out of two. *, unspecific band; black arrow, pT592-SAMHD1 specific band.

Techniques: Phospho-proteomics, Western Blot, Infection, Expressing, Control

a-h) HFFs were transfected with SAMHD1 siRNA or non-targeting siRNA (ctrl). Two days later, cells were either infected with AD169 at an MOI of 1 and harvested at 3 dpi (a-e) or infected at an MOI of 0.1 or 0.05 and harvested at 6 dpi (f-h). a) Levels of SAMHD1 and IE1/IE2 viral protein expression were assayed by immunoblotting. The p85 subunit of PI3K was used as loading control. One representative experiment is shown. b) The relative amount of SAMHD1 protein, normalized to that of p85, was determined by densitometric analysis and is relative to that of n.i./siCtrl cells, which was arbitrarily set as 1. Data are expressed as mean ± SE. c) The percentage of IE+ cells was analyzed by FACS after intracellular staining with a specific anti-IE1/IE2 mAb. FACS plots derive from one representative experiment. d) The percentage of IE+ cells is expressed as mean ± SD, as detected by FACS. e) Cell culture supernatants were assayed for infectious virus production by plaque assays. Results are expressed as mean ± SE. All data in panels a-e derive from five independent experiments. f) Immunoblotting at 6 dpi was performed as described in panel a). One representative experiment out of two is shown. g) The relative amount of SAMHD1 protein was determined as in panel b). Data are expressed as mean ± SE of two independent experiments with cells infected at both MOI. h) Viral titers were measured and expressed as in panel e) and derive from cell culture supernatants of two independent experiments, were HFFs were infected at an MOI of 0.1 or MOI of 0.05 for 6 days. i-j) HFFs were infected with single-cycle VLPs loaded with Vpx (Vpx) or unloaded (ΔVpx) at an MOI of 1, and then the same cells were infected with AD169 at an MOI of 1. At different dpi, cells and supernatants were harvested and subjected to immunoblotting and plaque assays, respectively. i) Immunoblotting of SAMHD1 protein expression. One representative experiment out of three is shown. j) Cell culture supernatants were assayed for infectious virus production by plaque assays. Results are expressed as mean ± SE of four experiments at 2–3 dpi. nt, not treated; n.i., not infected cells; *, p < 0.05; ***, p < 0.001; ns, not significant.

Journal: PLoS Pathogens

Article Title: SAMHD1 phosphorylation and cytoplasmic relocalization after human cytomegalovirus infection limits its antiviral activity

doi: 10.1371/journal.ppat.1008855

Figure Lengend Snippet: a-h) HFFs were transfected with SAMHD1 siRNA or non-targeting siRNA (ctrl). Two days later, cells were either infected with AD169 at an MOI of 1 and harvested at 3 dpi (a-e) or infected at an MOI of 0.1 or 0.05 and harvested at 6 dpi (f-h). a) Levels of SAMHD1 and IE1/IE2 viral protein expression were assayed by immunoblotting. The p85 subunit of PI3K was used as loading control. One representative experiment is shown. b) The relative amount of SAMHD1 protein, normalized to that of p85, was determined by densitometric analysis and is relative to that of n.i./siCtrl cells, which was arbitrarily set as 1. Data are expressed as mean ± SE. c) The percentage of IE+ cells was analyzed by FACS after intracellular staining with a specific anti-IE1/IE2 mAb. FACS plots derive from one representative experiment. d) The percentage of IE+ cells is expressed as mean ± SD, as detected by FACS. e) Cell culture supernatants were assayed for infectious virus production by plaque assays. Results are expressed as mean ± SE. All data in panels a-e derive from five independent experiments. f) Immunoblotting at 6 dpi was performed as described in panel a). One representative experiment out of two is shown. g) The relative amount of SAMHD1 protein was determined as in panel b). Data are expressed as mean ± SE of two independent experiments with cells infected at both MOI. h) Viral titers were measured and expressed as in panel e) and derive from cell culture supernatants of two independent experiments, were HFFs were infected at an MOI of 0.1 or MOI of 0.05 for 6 days. i-j) HFFs were infected with single-cycle VLPs loaded with Vpx (Vpx) or unloaded (ΔVpx) at an MOI of 1, and then the same cells were infected with AD169 at an MOI of 1. At different dpi, cells and supernatants were harvested and subjected to immunoblotting and plaque assays, respectively. i) Immunoblotting of SAMHD1 protein expression. One representative experiment out of three is shown. j) Cell culture supernatants were assayed for infectious virus production by plaque assays. Results are expressed as mean ± SE of four experiments at 2–3 dpi. nt, not treated; n.i., not infected cells; *, p < 0.05; ***, p < 0.001; ns, not significant.

Techniques: Transfection, Infection, Expressing, Western Blot, Control, Staining, Cell Culture, Virus

a) HFFs were not infected (n.i.) or infected with AD169 at the indicated MOI, in the presence of different concentrations of the pUL97 inhibitor Maribavir (MBV). After removing virus inoculum, MBV was newly added and kept until cell harvesting at 1 dpi. Total or pT592-SAMHD1 levels, expression of IE1/IE2 viral antigens and of p85 were analyzed by immunoblotting as above. A representative experiment out of two performed at 8 hpi, 1, 3 and 6 dpi is shown. b) The relative amount of pT592-SAMHD1 normalized to that of p85 was determined by densitometric analysis and is relative to that of DMSO-treated cells, which was arbitrarily set as 1. Data are expressed as mean ± SE of two independent experiments (MOI 1, 1 dpi). The analysis revealed no statistically significant difference. c-d) Effect of MBV on infectious virus production as measured by standard plaque assays on cell culture supernatants at 3 dpi (c) or 6 dpi) (d). Results derive from one representative experiment out of two. e) Total or pT592-SAMHD1 was analyzed as in panel a) in cells treated with the inhibitor Gö6976 (Gö). A representative experiment out of two performed at 3 dpi is shown. f) HFFs were transfected with three different UL97 siRNAs, alone (1, 2, 3) or in combination (1+2+3), or with a non-targeting siRNA (ctrl). The day after, cells were infected with AD169 at an MOI of 1 and harvested at 3 dpi. Immunoblotting was performed as above. One representative experiment out of two is shown. NT, not treated cells; DMSO, cells treated with the vehicle DMSO; *, unspecific band; black arrow, pT592-SAMHD1 specific band.

Journal: PLoS Pathogens

Article Title: SAMHD1 phosphorylation and cytoplasmic relocalization after human cytomegalovirus infection limits its antiviral activity

doi: 10.1371/journal.ppat.1008855

Figure Lengend Snippet: a) HFFs were not infected (n.i.) or infected with AD169 at the indicated MOI, in the presence of different concentrations of the pUL97 inhibitor Maribavir (MBV). After removing virus inoculum, MBV was newly added and kept until cell harvesting at 1 dpi. Total or pT592-SAMHD1 levels, expression of IE1/IE2 viral antigens and of p85 were analyzed by immunoblotting as above. A representative experiment out of two performed at 8 hpi, 1, 3 and 6 dpi is shown. b) The relative amount of pT592-SAMHD1 normalized to that of p85 was determined by densitometric analysis and is relative to that of DMSO-treated cells, which was arbitrarily set as 1. Data are expressed as mean ± SE of two independent experiments (MOI 1, 1 dpi). The analysis revealed no statistically significant difference. c-d) Effect of MBV on infectious virus production as measured by standard plaque assays on cell culture supernatants at 3 dpi (c) or 6 dpi) (d). Results derive from one representative experiment out of two. e) Total or pT592-SAMHD1 was analyzed as in panel a) in cells treated with the inhibitor Gö6976 (Gö). A representative experiment out of two performed at 3 dpi is shown. f) HFFs were transfected with three different UL97 siRNAs, alone (1, 2, 3) or in combination (1+2+3), or with a non-targeting siRNA (ctrl). The day after, cells were infected with AD169 at an MOI of 1 and harvested at 3 dpi. Immunoblotting was performed as above. One representative experiment out of two is shown. NT, not treated cells; DMSO, cells treated with the vehicle DMSO; *, unspecific band; black arrow, pT592-SAMHD1 specific band.

Techniques: Infection, Virus, Cell Harvesting, Expressing, Western Blot, Cell Culture, Transfection

a) HFFs were pretreated for 5 h with the Cdk1 inhibitor CGP74514A (CGP), and then not infected or infected with AD169, in the presence of CGP. After removing virus inoculum, the inhibitor was newly added and kept until cell harvesting, at 3 dpi. Total or pT592-SAMHD1 levels were analyzed by immunoblotting, together with IE1/IE2 and p85 expression. A representative experiment out of five is shown. b) The relative amount of pT592-SAMHD1, normalized to that of p85, was determined by densitometric analysis and is relative to that of n.i. cells, which was arbitrarily set as 1. Data are expressed as mean ± SE of four independent experiments performed at 1 and 3 dpi. c) Effect of CGP on infectious virus production as measured by standard plaque assays on cell culture supernatants at 3 dpi. Results derive from seven experiments and are expressed as mean ± SE. d) HFFs were pretreated as in panel a) with the Cdk inhibitors RO-3306 (RO), KO-3861 (KO) or BMS-265246 (BMS) at the indicated concentrations (μM), and then not infected or infected with AD169, in the presence of the inhibitors. After removing virus inoculum, the drugs were newly added and kept until cell harvesting, at 1 dpi. Immunoblotting was performed as above. One representative experiment out of two is shown. e) HFFs were transfected with Cdk1 or Cdk2 siRNA or non-targeting siRNA (ctrl). The day after, cells were not infected or infected with AD169 at an MOI of 1 and harvested at 1 dpi. Immunoblotting was performed as above. One representative experiment out of two is shown. NT, not treated cells; DMSO, cells treated with the vehicle DMSO; *, unspecific band; black arrow, pT592-SAMHD1 specific band. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; *****, p< 0.00001.

Journal: PLoS Pathogens

Article Title: SAMHD1 phosphorylation and cytoplasmic relocalization after human cytomegalovirus infection limits its antiviral activity

doi: 10.1371/journal.ppat.1008855

Figure Lengend Snippet: a) HFFs were pretreated for 5 h with the Cdk1 inhibitor CGP74514A (CGP), and then not infected or infected with AD169, in the presence of CGP. After removing virus inoculum, the inhibitor was newly added and kept until cell harvesting, at 3 dpi. Total or pT592-SAMHD1 levels were analyzed by immunoblotting, together with IE1/IE2 and p85 expression. A representative experiment out of five is shown. b) The relative amount of pT592-SAMHD1, normalized to that of p85, was determined by densitometric analysis and is relative to that of n.i. cells, which was arbitrarily set as 1. Data are expressed as mean ± SE of four independent experiments performed at 1 and 3 dpi. c) Effect of CGP on infectious virus production as measured by standard plaque assays on cell culture supernatants at 3 dpi. Results derive from seven experiments and are expressed as mean ± SE. d) HFFs were pretreated as in panel a) with the Cdk inhibitors RO-3306 (RO), KO-3861 (KO) or BMS-265246 (BMS) at the indicated concentrations (μM), and then not infected or infected with AD169, in the presence of the inhibitors. After removing virus inoculum, the drugs were newly added and kept until cell harvesting, at 1 dpi. Immunoblotting was performed as above. One representative experiment out of two is shown. e) HFFs were transfected with Cdk1 or Cdk2 siRNA or non-targeting siRNA (ctrl). The day after, cells were not infected or infected with AD169 at an MOI of 1 and harvested at 1 dpi. Immunoblotting was performed as above. One representative experiment out of two is shown. NT, not treated cells; DMSO, cells treated with the vehicle DMSO; *, unspecific band; black arrow, pT592-SAMHD1 specific band. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001; *****, p< 0.00001.

Techniques: Infection, Virus, Cell Harvesting, Western Blot, Expressing, Cell Culture, Transfection

HFFs were not infected (n.i.) or infected with AD169 at an MOI of 1 for 3 days. Stainings were then performed using primary antibodies directed against total (c and g) or pT592-SAMHD1 (m and q), followed by Alexa-Fluor 594 conjugated goat anti-rabbit (red), together with IE1/IE2 viral antigens followed by Alexa-Fluor 488 conjugated goat anti-mouse (a, e, k, o) (green) antibodies. Nuclei were stained with DAPI (b, f, l, p) (blue). Confocal images were acquired with a zoom 2 and are shown as single optical slice from one representative experiment out of six. An overlay of blue/red images (d, h, n, r) is also shown. Scale bar: 10μm. Fluorescence intensity (F.I.) of SAMHD1 (i-j) and p-T592 SAMHD1 staining (s-t) was calculated in the nucleus and cytoplasm of 80 cells randomly acquired from 3 independent experiments, as described in materials and methods. Histograms represent the mean ± SE. Displayed values are millions of a.u. ns, not statistically significant difference; ** p < 0.01; *** p < 0.001.

Journal: PLoS Pathogens

Article Title: SAMHD1 phosphorylation and cytoplasmic relocalization after human cytomegalovirus infection limits its antiviral activity

doi: 10.1371/journal.ppat.1008855

Figure Lengend Snippet: HFFs were not infected (n.i.) or infected with AD169 at an MOI of 1 for 3 days. Stainings were then performed using primary antibodies directed against total (c and g) or pT592-SAMHD1 (m and q), followed by Alexa-Fluor 594 conjugated goat anti-rabbit (red), together with IE1/IE2 viral antigens followed by Alexa-Fluor 488 conjugated goat anti-mouse (a, e, k, o) (green) antibodies. Nuclei were stained with DAPI (b, f, l, p) (blue). Confocal images were acquired with a zoom 2 and are shown as single optical slice from one representative experiment out of six. An overlay of blue/red images (d, h, n, r) is also shown. Scale bar: 10μm. Fluorescence intensity (F.I.) of SAMHD1 (i-j) and p-T592 SAMHD1 staining (s-t) was calculated in the nucleus and cytoplasm of 80 cells randomly acquired from 3 independent experiments, as described in materials and methods. Histograms represent the mean ± SE. Displayed values are millions of a.u. ns, not statistically significant difference; ** p < 0.01; *** p < 0.001.

Techniques: Infection, Staining, Fluorescence

$a) HFFs were infected with HCMV at an MOI of 1 and nuclear, cytoplasmic, and total cell extracts were prepared at 3 dpi. Thirty μg of proteins were loaded and subjected to immunoblotting with the indicated antibodies. Lamin-A and α-tubulin were used to confirm fractionation efficiency, and p85 as loading control. Results derive from one representative experiment out of four. b-c) The relative amount of total (b) or pT592-SAMHD1 (c), normalized to that of p85, was determined by densitometric analysis and is relative to that of total cell extracts in n.i. cells, which was arbitrarily set as 1. Data are expressed as mean ± SE of four independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.$

Journal: PLoS Pathogens

Article Title: SAMHD1 phosphorylation and cytoplasmic relocalization after human cytomegalovirus infection limits its antiviral activity

doi: 10.1371/journal.ppat.1008855

Figure Lengend Snippet: a) HFFs were infected with HCMV at an MOI of 1 and nuclear, cytoplasmic, and total cell extracts were prepared at 3 dpi. Thirty μg of proteins were loaded and subjected to immunoblotting with the indicated antibodies. Lamin-A and α-tubulin were used to confirm fractionation efficiency, and p85 as loading control. Results derive from one representative experiment out of four. b-c) The relative amount of total (b) or pT592-SAMHD1 (c), normalized to that of p85, was determined by densitometric analysis and is relative to that of total cell extracts in n.i. cells, which was arbitrarily set as 1. Data are expressed as mean ± SE of four independent experiments. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ****, p < 0.0001.

Techniques: Infection, Western Blot, Fractionation, Control

HFFs, not infected (n.i.) or infected with AD169, were processed at 3 dpi for Cryo-IEM analysis and immunolabeled using primary specific antibodies against SAMHD1 (a-d) or pT592-SAMHD1 (e-h). One representative experiment out of seven different immune gold-labeling experiments is shown. Sparse labeling of SAMHD1 and pT592-SAMHD1 was detected in uninfected cells in the nucleus and in the cytoplasm (a, e). In infected cells, viral structures were not labeled with SAMHD1 gold (b-d; white arrows) but were gold-decorated with pT592-SAMHD1 (f-h; black arrows). N, nucleus; Ne, nuclear envelope; G, Golgi apparatus, DB, dense bodies. Bars a, b, d, e, g, h = 200nm; c, f = 100 nm.

Journal: PLoS Pathogens

Article Title: SAMHD1 phosphorylation and cytoplasmic relocalization after human cytomegalovirus infection limits its antiviral activity

doi: 10.1371/journal.ppat.1008855

Figure Lengend Snippet: HFFs, not infected (n.i.) or infected with AD169, were processed at 3 dpi for Cryo-IEM analysis and immunolabeled using primary specific antibodies against SAMHD1 (a-d) or pT592-SAMHD1 (e-h). One representative experiment out of seven different immune gold-labeling experiments is shown. Sparse labeling of SAMHD1 and pT592-SAMHD1 was detected in uninfected cells in the nucleus and in the cytoplasm (a, e). In infected cells, viral structures were not labeled with SAMHD1 gold (b-d; white arrows) but were gold-decorated with pT592-SAMHD1 (f-h; black arrows). N, nucleus; Ne, nuclear envelope; G, Golgi apparatus, DB, dense bodies. Bars a, b, d, e, g, h = 200nm; c, f = 100 nm.

Techniques: Infection, Immunolabeling, Labeling

Immunogold quantification of total and pT592-SAMHD1 on virions and dense bodies in infected cells (MOI 1).

Journal: PLoS Pathogens

Article Title: SAMHD1 phosphorylation and cytoplasmic relocalization after human cytomegalovirus infection limits its antiviral activity

doi: 10.1371/journal.ppat.1008855

Figure Lengend Snippet: Immunogold quantification of total and pT592-SAMHD1 on virions and dense bodies in infected cells (MOI 1).

Techniques: Infection

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