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rabbit polyclonal antibody against ddx6 (Cell Signaling Technology Inc)

Name: rabbit polyclonal antibody against ddx6
Brand: Cell Signaling Technology Inc
Rating: 93 (23 reviews)

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Cell Signaling Technology Inc rabbit polyclonal antibody against ddx6

Fig. 2. Various PB markers accumulated in the HFF cells infected with HCMV. (A) The amount of different PB markers increased during HCMV infection. HFF cells were infected with HCMV at an moi of 3 in the absence or presence of 200 μM cycloheximide and then harvested at 0, 6, 24, and 48 hpi in the absence of cycloheximide and 0 and 24 hpi in the presence of 200 μM cycloheximide. Immunoblot analysis was performed using an antibody against HCMV IE1 (IE72) and IE2 (IE86), DCP1a, EDC4, <t>DDX6,</t> 4E T, Lsm1, Lsm14A, or GAPDH at the times indicated after HCMV infection. GAPDH served as a loading control. (B) Accumulation of various PB markers during HCMV infection. HFF cells were treated with 0.5 mM Arsenite for 30 min or infected with HCMV at an moi of 3 in the absence or presence of 200 μM cycloheximide and ﬁxed at 0 and 24 hpi. The cells were stained with mouse monoclonal or rabbit <t>polyclonal</t> antibody speciﬁc for Dcp1a or Lsm14A, respectively, and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 568-conjugated donkey anti- mouse or anti-rabbit IgG, respectively, and Alexa 680-conjugated anti-goat secondary antibodies. Histogram shows the ratio of PBs/cell counted using Dcp1a and EDC4 PB markers. Left and right bars represent the number of PBs with a diameter of less or more than 1 μm, respectively. nn indicates values of po0.01. (C) HFF cells were infected with HCMV at an moi of 3, ﬁxed at 0 and 24 hpi, and then stained with mouse monoclonal antibody speciﬁc for Ago2 and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 568- conjugated donkey anti-mouse IgG and Alexa 680-conjugated anti-goat secondary antibodies.

Rabbit Polyclonal Antibody Against Ddx6, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 93 stars, based on 23 article reviews

rabbit polyclonal antibody against ddx6 - by Bioz Stars, 2026-02

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1) Product Images from "Processing bodies accumulate in human cytomegalovirus-infected cells and do not affect viral replication at high multiplicity of infection."

Article Title: Processing bodies accumulate in human cytomegalovirus-infected cells and do not affect viral replication at high multiplicity of infection.

Journal: Virology

doi: 10.1016/j.virol.2014.04.022

Figure Legend Snippet: Fig. 2. Various PB markers accumulated in the HFF cells infected with HCMV. (A) The amount of different PB markers increased during HCMV infection. HFF cells were infected with HCMV at an moi of 3 in the absence or presence of 200 μM cycloheximide and then harvested at 0, 6, 24, and 48 hpi in the absence of cycloheximide and 0 and 24 hpi in the presence of 200 μM cycloheximide. Immunoblot analysis was performed using an antibody against HCMV IE1 (IE72) and IE2 (IE86), DCP1a, EDC4, DDX6, 4E T, Lsm1, Lsm14A, or GAPDH at the times indicated after HCMV infection. GAPDH served as a loading control. (B) Accumulation of various PB markers during HCMV infection. HFF cells were treated with 0.5 mM Arsenite for 30 min or infected with HCMV at an moi of 3 in the absence or presence of 200 μM cycloheximide and ﬁxed at 0 and 24 hpi. The cells were stained with mouse monoclonal or rabbit polyclonal antibody speciﬁc for Dcp1a or Lsm14A, respectively, and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 568-conjugated donkey anti- mouse or anti-rabbit IgG, respectively, and Alexa 680-conjugated anti-goat secondary antibodies. Histogram shows the ratio of PBs/cell counted using Dcp1a and EDC4 PB markers. Left and right bars represent the number of PBs with a diameter of less or more than 1 μm, respectively. nn indicates values of po0.01. (C) HFF cells were infected with HCMV at an moi of 3, ﬁxed at 0 and 24 hpi, and then stained with mouse monoclonal antibody speciﬁc for Ago2 and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 568- conjugated donkey anti-mouse IgG and Alexa 680-conjugated anti-goat secondary antibodies.

Techniques Used: Infection, Western Blot, Control, Staining

Fig. 4. PB accumulation in HFF cells infected with HCMV or UV-inactivated HCMV. HFF cells were infected with HCMV (A or C), or UV-inactivated virions (B and D), or the culture ﬂuid after removal of HCMV virions (E) at an moi of 3. HFF cells were maintained in the absence (A and B) or presence (C and D) of cycloheximide. Cells were analyzed for IE1 RNA by FISH and then stained with rabbit polyclonal antibody speciﬁc for Lsm14A and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 488-conjugated donkey anti-rabbit and Alexa 680-conjugated anti-goat IgG secondary antibodies. (F) Histogram showing the number of PBs/cell. Thirty cells were selected from multiple ﬁelds of view, and the number of Lsm14A and EDC4 double-positive PBs were counted. Data are averages of merged dots determined by counting three times independently, and the error bars indicate SD.

Figure Legend Snippet: Fig. 4. PB accumulation in HFF cells infected with HCMV or UV-inactivated HCMV. HFF cells were infected with HCMV (A or C), or UV-inactivated virions (B and D), or the culture ﬂuid after removal of HCMV virions (E) at an moi of 3. HFF cells were maintained in the absence (A and B) or presence (C and D) of cycloheximide. Cells were analyzed for IE1 RNA by FISH and then stained with rabbit polyclonal antibody speciﬁc for Lsm14A and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 488-conjugated donkey anti-rabbit and Alexa 680-conjugated anti-goat IgG secondary antibodies. (F) Histogram showing the number of PBs/cell. Thirty cells were selected from multiple ﬁelds of view, and the number of Lsm14A and EDC4 double-positive PBs were counted. Data are averages of merged dots determined by counting three times independently, and the error bars indicate SD.

Techniques Used: Infection, Staining

Fig. 5. Cytoplasmic distributions of IE1 mRNA and PB markers, Lsm14A and EDC4. HFF cells were infected with HCMV at an moi of 3 and ﬁxed at 1 (A), 3 (B), 6 (C), and 24 (D) hpi. Infected HFF cells were also maintained in the presence of cycloheximide and ﬁxed at 24 hpi (E). These cells were analyzed with IE1 probe by FISH and then stained with rabbit polyclonal antibody speciﬁc for Lsm14A and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 488-conjugated donkey anti-rabbit IgG and Alexa 680-conjugated anti-goat secondary antibodies.

Figure Legend Snippet: Fig. 5. Cytoplasmic distributions of IE1 mRNA and PB markers, Lsm14A and EDC4. HFF cells were infected with HCMV at an moi of 3 and ﬁxed at 1 (A), 3 (B), 6 (C), and 24 (D) hpi. Infected HFF cells were also maintained in the presence of cycloheximide and ﬁxed at 24 hpi (E). These cells were analyzed with IE1 probe by FISH and then stained with rabbit polyclonal antibody speciﬁc for Lsm14A and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 488-conjugated donkey anti-rabbit IgG and Alexa 680-conjugated anti-goat secondary antibodies.

Techniques Used: Infection, Staining

Fig. 6. Knockdown effect of PB components on viral gene expression in HCMV-infected cells. HFF cells were transfected with siRNAs targeting 4E-T, Lsm1, and LSM14A, or with a non-targeting siRNA (siNeg), and, 2 days after transfection, infected with HCMV at an moi of 3. (A) Effect of siRNAs speciﬁc for the PB components, 4E-T, Lsm1, and Lsm14A on mRNA expression of the corresponding gene and viral mRNA expression. The transfected and infected cells were harvested at 0, 3, 6, and 24 hpi. Total RNA was isolated and subjected to quantitative real-time RT-PCR analysis to detect the gene transcripts indicated on the Y axis. RNAs were normalized to HPRT RNA and values were calculated relative to siNeg at 0 (upper panel) or 3 (lower panel) hpi. Data are averages of three independent experiments. Statistical analyses were done using the t-test. RNA expression levels compared to siNeg were assessed for statistical signiﬁcance. n indicates values of po0.05. nn indicates values of po0.01. (B) The transfected and infected cells were ﬁxed at 24 hpi and stained with rabbit polyclonal antibody speciﬁc for Lsm14A and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 568-conjugated donkey anti-rabbit IgG and Alexa 680-conjugated anti-goat secondary antibodies. Histogram showing the effect of siRNAs targeting 4E-T, Lsm1, and LSM14A, or with a non-targeting siRNA (siNeg) on the ratio of the PB number/cell. Thirty cells were selected from multiple ﬁelds of view, and the number of Lsm14A and EDC4 double- positive PBs were counted. Data are averages of merged dots determined by counting three times independently, and the error bars indicate SD.

Figure Legend Snippet: Fig. 6. Knockdown effect of PB components on viral gene expression in HCMV-infected cells. HFF cells were transfected with siRNAs targeting 4E-T, Lsm1, and LSM14A, or with a non-targeting siRNA (siNeg), and, 2 days after transfection, infected with HCMV at an moi of 3. (A) Effect of siRNAs speciﬁc for the PB components, 4E-T, Lsm1, and Lsm14A on mRNA expression of the corresponding gene and viral mRNA expression. The transfected and infected cells were harvested at 0, 3, 6, and 24 hpi. Total RNA was isolated and subjected to quantitative real-time RT-PCR analysis to detect the gene transcripts indicated on the Y axis. RNAs were normalized to HPRT RNA and values were calculated relative to siNeg at 0 (upper panel) or 3 (lower panel) hpi. Data are averages of three independent experiments. Statistical analyses were done using the t-test. RNA expression levels compared to siNeg were assessed for statistical signiﬁcance. n indicates values of po0.05. nn indicates values of po0.01. (B) The transfected and infected cells were ﬁxed at 24 hpi and stained with rabbit polyclonal antibody speciﬁc for Lsm14A and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 568-conjugated donkey anti-rabbit IgG and Alexa 680-conjugated anti-goat secondary antibodies. Histogram showing the effect of siRNAs targeting 4E-T, Lsm1, and LSM14A, or with a non-targeting siRNA (siNeg) on the ratio of the PB number/cell. Thirty cells were selected from multiple ﬁelds of view, and the number of Lsm14A and EDC4 double- positive PBs were counted. Data are averages of merged dots determined by counting three times independently, and the error bars indicate SD.

Techniques Used: Knockdown, Gene Expression, Infection, Transfection, Expressing, Isolation, Quantitative RT-PCR, RNA Expression, Staining

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Journal: Virology

Article Title: Processing bodies accumulate in human cytomegalovirus-infected cells and do not affect viral replication at high multiplicity of infection.

doi: 10.1016/j.virol.2014.04.022

Figure Lengend Snippet: Fig. 2. Various PB markers accumulated in the HFF cells infected with HCMV. (A) The amount of different PB markers increased during HCMV infection. HFF cells were infected with HCMV at an moi of 3 in the absence or presence of 200 μM cycloheximide and then harvested at 0, 6, 24, and 48 hpi in the absence of cycloheximide and 0 and 24 hpi in the presence of 200 μM cycloheximide. Immunoblot analysis was performed using an antibody against HCMV IE1 (IE72) and IE2 (IE86), DCP1a, EDC4, DDX6, 4E T, Lsm1, Lsm14A, or GAPDH at the times indicated after HCMV infection. GAPDH served as a loading control. (B) Accumulation of various PB markers during HCMV infection. HFF cells were treated with 0.5 mM Arsenite for 30 min or infected with HCMV at an moi of 3 in the absence or presence of 200 μM cycloheximide and ﬁxed at 0 and 24 hpi. The cells were stained with mouse monoclonal or rabbit polyclonal antibody speciﬁc for Dcp1a or Lsm14A, respectively, and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 568-conjugated donkey anti- mouse or anti-rabbit IgG, respectively, and Alexa 680-conjugated anti-goat secondary antibodies. Histogram shows the ratio of PBs/cell counted using Dcp1a and EDC4 PB markers. Left and right bars represent the number of PBs with a diameter of less or more than 1 μm, respectively. nn indicates values of po0.01. (C) HFF cells were infected with HCMV at an moi of 3, ﬁxed at 0 and 24 hpi, and then stained with mouse monoclonal antibody speciﬁc for Ago2 and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 568- conjugated donkey anti-mouse IgG and Alexa 680-conjugated anti-goat secondary antibodies.

Article Snippet: Rabbit polyclonal antibody against DDX6 (Cat# 9407) was from Cell Signaling (Beverly, MA) and rabbit polyclonal antibody against Lsm14A (Cat# ABE37) was from Millipore (Billerica, MA).

Techniques: Infection, Western Blot, Control, Staining

Journal: Virology

Article Title: Processing bodies accumulate in human cytomegalovirus-infected cells and do not affect viral replication at high multiplicity of infection.

doi: 10.1016/j.virol.2014.04.022

Figure Lengend Snippet: Fig. 4. PB accumulation in HFF cells infected with HCMV or UV-inactivated HCMV. HFF cells were infected with HCMV (A or C), or UV-inactivated virions (B and D), or the culture ﬂuid after removal of HCMV virions (E) at an moi of 3. HFF cells were maintained in the absence (A and B) or presence (C and D) of cycloheximide. Cells were analyzed for IE1 RNA by FISH and then stained with rabbit polyclonal antibody speciﬁc for Lsm14A and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 488-conjugated donkey anti-rabbit and Alexa 680-conjugated anti-goat IgG secondary antibodies. (F) Histogram showing the number of PBs/cell. Thirty cells were selected from multiple ﬁelds of view, and the number of Lsm14A and EDC4 double-positive PBs were counted. Data are averages of merged dots determined by counting three times independently, and the error bars indicate SD.

Techniques: Infection, Staining

Journal: Virology

Article Title: Processing bodies accumulate in human cytomegalovirus-infected cells and do not affect viral replication at high multiplicity of infection.

doi: 10.1016/j.virol.2014.04.022

Figure Lengend Snippet: Fig. 5. Cytoplasmic distributions of IE1 mRNA and PB markers, Lsm14A and EDC4. HFF cells were infected with HCMV at an moi of 3 and ﬁxed at 1 (A), 3 (B), 6 (C), and 24 (D) hpi. Infected HFF cells were also maintained in the presence of cycloheximide and ﬁxed at 24 hpi (E). These cells were analyzed with IE1 probe by FISH and then stained with rabbit polyclonal antibody speciﬁc for Lsm14A and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 488-conjugated donkey anti-rabbit IgG and Alexa 680-conjugated anti-goat secondary antibodies.

Techniques: Infection, Staining

Journal: Virology

Article Title: Processing bodies accumulate in human cytomegalovirus-infected cells and do not affect viral replication at high multiplicity of infection.

doi: 10.1016/j.virol.2014.04.022

Figure Lengend Snippet: Fig. 6. Knockdown effect of PB components on viral gene expression in HCMV-infected cells. HFF cells were transfected with siRNAs targeting 4E-T, Lsm1, and LSM14A, or with a non-targeting siRNA (siNeg), and, 2 days after transfection, infected with HCMV at an moi of 3. (A) Effect of siRNAs speciﬁc for the PB components, 4E-T, Lsm1, and Lsm14A on mRNA expression of the corresponding gene and viral mRNA expression. The transfected and infected cells were harvested at 0, 3, 6, and 24 hpi. Total RNA was isolated and subjected to quantitative real-time RT-PCR analysis to detect the gene transcripts indicated on the Y axis. RNAs were normalized to HPRT RNA and values were calculated relative to siNeg at 0 (upper panel) or 3 (lower panel) hpi. Data are averages of three independent experiments. Statistical analyses were done using the t-test. RNA expression levels compared to siNeg were assessed for statistical signiﬁcance. n indicates values of po0.05. nn indicates values of po0.01. (B) The transfected and infected cells were ﬁxed at 24 hpi and stained with rabbit polyclonal antibody speciﬁc for Lsm14A and goat polyclonal antibody speciﬁc for EDC4, followed by staining with Alexa 568-conjugated donkey anti-rabbit IgG and Alexa 680-conjugated anti-goat secondary antibodies. Histogram showing the effect of siRNAs targeting 4E-T, Lsm1, and LSM14A, or with a non-targeting siRNA (siNeg) on the ratio of the PB number/cell. Thirty cells were selected from multiple ﬁelds of view, and the number of Lsm14A and EDC4 double- positive PBs were counted. Data are averages of merged dots determined by counting three times independently, and the error bars indicate SD.

Techniques: Knockdown, Gene Expression, Infection, Transfection, Expressing, Isolation, Quantitative RT-PCR, RNA Expression, Staining

$Pat1b interacts with P-body proteins. (A) HEK293 cells were transiently transfected with vector alone, HA-tagged Pat1b, or HA-tagged Pat1a. After 1 day, cytoplasmic lysates (input) were prepared for IP with anti-HA antibody. The HA-tagged proteins as well as endogenous Rck, Hedls, Xrn1, Lsm1, Lsm4, and eIF3B were detected by Western blotting. The sizes of the molecular weight markers (in thousands) are indicated on the right. (B) HEK293 cells transiently transfected with vector alone, HA-Pat1b, or HA-Pat1b-YFP were used for IP as described in the legend for panel A. Where indicated, RNase A was added to the lysates during IP. Lanes 10 and 11 show RNA extracted from the unbound fraction and stained with ethidium bromide. (C) HA-Pat1b was immunoprecipitated with HA antibody or without antibody as a control for unspecific precipitation. (D) HA-Pat1b was immunoprecipitated and subjected to increasing NaCl concentrations prior to elution of the protein complexes. *, immunoglobulin heavy chain. (E) Endogenous Xrn1, Hedls, Rck, Lsm1, and 14-3-3 were immunoprecipitated from the cytoplasmic lysate of HEK293 cells transiently transfected with HA-Pat1b. The Western blot for Rck is not shown due to an overlapping signal from the immunoglobulin heavy chain.$

Journal: Molecular and Cellular Biology

Article Title: Human Pat1b Connects Deadenylation with mRNA Decapping and Controls the Assembly of Processing Bodies ▿ †

doi: 10.1128/MCB.00429-10

Figure Lengend Snippet: Pat1b interacts with P-body proteins. (A) HEK293 cells were transiently transfected with vector alone, HA-tagged Pat1b, or HA-tagged Pat1a. After 1 day, cytoplasmic lysates (input) were prepared for IP with anti-HA antibody. The HA-tagged proteins as well as endogenous Rck, Hedls, Xrn1, Lsm1, Lsm4, and eIF3B were detected by Western blotting. The sizes of the molecular weight markers (in thousands) are indicated on the right. (B) HEK293 cells transiently transfected with vector alone, HA-Pat1b, or HA-Pat1b-YFP were used for IP as described in the legend for panel A. Where indicated, RNase A was added to the lysates during IP. Lanes 10 and 11 show RNA extracted from the unbound fraction and stained with ethidium bromide. (C) HA-Pat1b was immunoprecipitated with HA antibody or without antibody as a control for unspecific precipitation. (D) HA-Pat1b was immunoprecipitated and subjected to increasing NaCl concentrations prior to elution of the protein complexes. *, immunoglobulin heavy chain. (E) Endogenous Xrn1, Hedls, Rck, Lsm1, and 14-3-3 were immunoprecipitated from the cytoplasmic lysate of HEK293 cells transiently transfected with HA-Pat1b. The Western blot for Rck is not shown due to an overlapping signal from the immunoglobulin heavy chain.

Article Snippet: The following antibodies were used for IP and Western blotting, and immunofluorescence analysis: mouse monoclonal antibodies anti-HA (HA.11; Covance) and anti-Edc3 (Abcam; ab57780-100); a cross-reacting mouse monoclonal phospho-S6-kinase antibody (sc-8416; Santa Cruz) for detection of Hedls; polyclonal chicken antibodies against Lsm1 (15-288-22100F; Genway Biotech) and Lsm4 (19101; Abcam); polyclonal rabbit antibodies against Xrn1 (A300-461A; Bethyl Laboratories), p54/Rck (A300-443A; Bethyl Laboratories), eIF4G (sc-11373; Santa Cruz), 14-3-3 (sc-629; Santa Cruz), and HA (sc-805; Santa Cruz); and a polyclonal goat antibody against eIF3B (sc-16377; Santa Cruz).

Techniques: Transfection, Plasmid Preparation, Western Blot, Molecular Weight, Staining, Immunoprecipitation, Control

Role of Pat1b in P-body assembly and mRNA degradation. (A) Schematic representation human Pat1b subdomains, together with interacting proteins and associated functions. All the interactions depicted are RNA independent yet may be direct or indirect. (B) Hypothetical model of the Pat1b-associated complex that connects deadenylation with mRNA decapping. The acidic region (A) associates with Rck, the AN fragment with the Ccr4-Not complex, and the N fragment with Caf1 and Dcp2-Dcp1a. The homology region (H) is required for Lsm1 binding, and the HC fragment further associates with Dcp2-Dcp1a. The HC fragment also shows weak association with Edc3, Hedls, and Xrn1.

Journal: Molecular and Cellular Biology

Article Title: Human Pat1b Connects Deadenylation with mRNA Decapping and Controls the Assembly of Processing Bodies ▿ †

doi: 10.1128/MCB.00429-10

Figure Lengend Snippet: Role of Pat1b in P-body assembly and mRNA degradation. (A) Schematic representation human Pat1b subdomains, together with interacting proteins and associated functions. All the interactions depicted are RNA independent yet may be direct or indirect. (B) Hypothetical model of the Pat1b-associated complex that connects deadenylation with mRNA decapping. The acidic region (A) associates with Rck, the AN fragment with the Ccr4-Not complex, and the N fragment with Caf1 and Dcp2-Dcp1a. The homology region (H) is required for Lsm1 binding, and the HC fragment further associates with Dcp2-Dcp1a. The HC fragment also shows weak association with Edc3, Hedls, and Xrn1.

Techniques: Binding Assay

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