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Proteintech ruvbl1
Ruvbl1, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 59 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/ruvbl1/RUVBL1+Antibody/pm41787535-155-7-8
Average 93 stars, based on 59 article reviews
ruvbl1 - by Bioz Stars, 2026-10
93/100 stars

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Related Articles

Immunohistochemistry:

Article Title: The R2TP complex stabilises E7 to drive human papillomavirus-mediated pathogenesis in cellular models of cervical cancer
Article Snippet: For the proximity ligation assay, the following antibodies were used: PIH1D1 (Thermo Fisher, Cat. PA5-61482), RPAP3 (Thermo Fisher, Cat. PA5-58334), Human Retinoblastoma Protein [Clone G3-245 (RUO), BD PharmingenTM, Cat. 554136], and HPV16 E7 (ED17) (Santa Cruz, Cat. sc-6981). .. For immunohistochemistry, the following antibodies were used: RUVBL1 (Proteintech, Cat. 10210-2-AP), PIH1D1 (Thermo Fisher, Cat. PA5-61482), and RPAP3 (Thermo Fisher, Cat. PA5-58334). .. All data analyses were done with GraphPad Prism 8.

Article Title: Hippocalcin-Like 1 blunts liver lipid metabolism to suppress tumorigenesis via directly targeting RUVBL1-mTOR signaling
Article Snippet: .. For IHC staining, liver sections were preincubated in normal goat serum for 15 min, and were incubated with anti-phos-mTOR (Cell signaling, #2976,1: 50), anti-phos-4EBP1 (Cell signaling, #2855, 1:100), anti-HPCAL1 (Sigma, SAB1307075; 1:200), anti-SCD1 (Abclonal, A16429; 1:400), anti-ACSS2 (Proteintech, 16087-1-AP, 1:400), RUVBL1 (Proteintech,10210-2-AP,1:400) and Ki-67 (Proteintech, 27309-1-AP, 1:5000) at room temperature. ..

Article Title: The R2TP complex stabilises E7 to drive human papillomavirus-mediated pathogenesis in cellular models of cervical cancer.
Article Snippet: For the proximity ligation assay, the following antibodies were used: PIH1D1 (Thermo Fisher, Cat. PA5-61482), RPAP3 (Thermo Fisher, Cat. PA5-58334), Human Retinoblastoma Protein [Clone G3-245 (RUO), BD PharmingenTM, Cat. 554136], and HPV16 E7 (ED17) (Santa Cruz, Cat. sc-6981). .. For immunohistochemistry, the following antibodies were used: RUVBL1 (Proteintech, Cat. 10210-2-AP), PIH1D1 (Thermo Fisher, Cat. PA5-61482), and RPAP3 (Thermo Fisher, Cat. PA5-58334). .. All data analyses were done with GraphPad Prism 8.

Blocking Assay:

Article Title: Systematic analysis of various RNA transcripts and construction of biological regulatory networks at the post-transcriptional level for chronic obstructive pulmonary disease
Article Snippet: Equal amounts of proteins were separated on SDS–polyacrylamide gel electrophoresis (PAGE) and transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA). .. After blocking with 5% skim milk for 1.5 h at room temperature, the membranes were incubated with primary antibodies against DDB2 (Proteintech, 10431-1-AP), RUVBL1 (Proteintech, 10210-2-AP), and GAPDH (Abcam, ab181602) overnight at 4 °C, followed by incubation with HRP-conjugated goat anti-rabbit secondary antibodies (ABclonal, AS014) for 1.5 h at room temperature. ..

Article Title: Systematic analysis of various RNA transcripts and construction of biological regulatory networks at the post-transcriptional level for chronic obstructive pulmonary disease.
Article Snippet: Equal amounts of proteins were separated on SDS–polyacrylamide gel electrophoresis (PAGE) and transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA). .. After blocking with 5% skim milk for 1.5 h at room temperature, the membranes were incubated with primary antibodies against DDB2 (Proteintech, 10431-1- AP), RUVBL1 (Proteintech, 10210-2-AP), and GAPDH (Abcam, ab181602) overnight at 4 °C, followed by incubation with HRP-conjugated goat anti-rabbit secondary antibodies (ABclonal, AS014) for 1.5 h at room temperature. ..

Incubation:

Article Title: Systematic analysis of various RNA transcripts and construction of biological regulatory networks at the post-transcriptional level for chronic obstructive pulmonary disease
Article Snippet: Equal amounts of proteins were separated on SDS–polyacrylamide gel electrophoresis (PAGE) and transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA). .. After blocking with 5% skim milk for 1.5 h at room temperature, the membranes were incubated with primary antibodies against DDB2 (Proteintech, 10431-1-AP), RUVBL1 (Proteintech, 10210-2-AP), and GAPDH (Abcam, ab181602) overnight at 4 °C, followed by incubation with HRP-conjugated goat anti-rabbit secondary antibodies (ABclonal, AS014) for 1.5 h at room temperature. ..

Article Title: Hippocalcin-Like 1 blunts liver lipid metabolism to suppress tumorigenesis via directly targeting RUVBL1-mTOR signaling
Article Snippet: .. For IHC staining, liver sections were preincubated in normal goat serum for 15 min, and were incubated with anti-phos-mTOR (Cell signaling, #2976,1: 50), anti-phos-4EBP1 (Cell signaling, #2855, 1:100), anti-HPCAL1 (Sigma, SAB1307075; 1:200), anti-SCD1 (Abclonal, A16429; 1:400), anti-ACSS2 (Proteintech, 16087-1-AP, 1:400), RUVBL1 (Proteintech,10210-2-AP,1:400) and Ki-67 (Proteintech, 27309-1-AP, 1:5000) at room temperature. ..

Article Title: Systematic analysis of various RNA transcripts and construction of biological regulatory networks at the post-transcriptional level for chronic obstructive pulmonary disease.
Article Snippet: Equal amounts of proteins were separated on SDS–polyacrylamide gel electrophoresis (PAGE) and transferred onto polyvinylidene fluoride (PVDF) membranes (Millipore, USA). .. After blocking with 5% skim milk for 1.5 h at room temperature, the membranes were incubated with primary antibodies against DDB2 (Proteintech, 10431-1- AP), RUVBL1 (Proteintech, 10210-2-AP), and GAPDH (Abcam, ab181602) overnight at 4 °C, followed by incubation with HRP-conjugated goat anti-rabbit secondary antibodies (ABclonal, AS014) for 1.5 h at room temperature. ..

Article Title: RUVBL1 promotes enzalutamide resistance of prostate tumors through the PLXNA1-CRAF-MAPK pathway.
Article Snippet: Although enzalutamide improves the overall survival of patients with metastatic prostate cancers, enzalutamide resistance (ENZR) will be inevitably developed.. Emerging evidence support that alternative oncogenic pathways may bypass the androgen receptor (AR) signaling to promote ENZR progression, however, the underpinning mechanisms remain poorly defined.. Here, we report that the expression of RuvB like AAA ATPase 1 (RUVBL1) is upregulated in ENZR cells and xenograft models and prostate tumors in patients.

other:

Article Title: Jaceosidin overcomes osimertinib resistance in lung cancer by inducing G2/M cycle arrest through targeting DDB1.
Article Snippet: This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record.. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article.. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.

Staining:

Article Title: Hippocalcin-Like 1 blunts liver lipid metabolism to suppress tumorigenesis via directly targeting RUVBL1-mTOR signaling
Article Snippet: .. For IHC staining, liver sections were preincubated in normal goat serum for 15 min, and were incubated with anti-phos-mTOR (Cell signaling, #2976,1: 50), anti-phos-4EBP1 (Cell signaling, #2855, 1:100), anti-HPCAL1 (Sigma, SAB1307075; 1:200), anti-SCD1 (Abclonal, A16429; 1:400), anti-ACSS2 (Proteintech, 16087-1-AP, 1:400), RUVBL1 (Proteintech,10210-2-AP,1:400) and Ki-67 (Proteintech, 27309-1-AP, 1:5000) at room temperature. ..

Western Blot:

Article Title: Hippocalcin-Like 1 blunts liver lipid metabolism to suppress tumorigenesis via directly targeting RUVBL1-mTOR signaling
Article Snippet: AZD-8055 was purchased from MedChemExpress (Shanghai, China). .. Antibodies for western blotting against HPCAL1, ACSS2, RUVBL1, GAPDH, his tag and HA tag were obtained from Proteintech (Wuhan, China). .. Antibody against phos-mTOR, and phos-4EBP1 and phos-4EBP1 were purchased from Cell Signaling Technology (USA).



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Eurofins single sirna targeting ruvbl1
(A) Workflow for the identification of RV-A16 2C interactors and their functional validation. HeLa-H1 cells were infected in triplicate with RV-A16 (MOI 20) for 4.5 h, 6 h or were left uninfected. Protein-protein interactions were then cross-linked with formaldehyde and cell lysates were immunoprecipitated using RV-A16 2C-specific or control antisera (2C-IP or control-IP). Eluates were TMT-labelled, multiplexed and analysed by LC-MS/MS. Differences between infected and uninfected conditions were assessed using two-sample Student’s t -test. Cellular proteins significantly enriched in infected 2C-IP fractions were further investigated for their role in RV-A16 replication through siRNA screening. The two most significant non-cytotoxic hits from the siRNA screen, <t>RUVBL1</t> and RUVBL2, were further validated using a small molecule inhibitor. (B-C) Proteomics analysis of 2C-IPs. RV-A16 2C was immunoprecipitated from infected cells and the associated proteins were analysed as described in A. (B) Volcano plots showing in red the cellular proteins significantly enriched in 2C-IPs at 4.5 hpi (top) and 6 hpi (bottom), compared to uninfected conditions. (C) Corresponding heatmap, also showing infected control-IPs. (D-E) siRNA screen of the 2C-IP hits. HeLa-H1 cells were transfected with siRNA pools targeting the 2C-IPs hits or a non-targeting (NT) control siRNA pool. (D) At 72 h post-transfection, cells were infected with RV-A16 (MOI 20), and viral titres were quantified at 6 hpi. Data are presented as a percentage of the mean NT control (dashed grey line). (E) Viability of uninfected cells at 72 h post-transfection, for siRNA significantly reducing viral replication, presented as percentage of the mean NT control (dashed grey line). Statistical tests: one-way ANOVA with Dunnett’s post-hoc test, comparing to the NT control pool. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (F-G) Validation using CB-6644, a small molecule inhibitor of the ATPase activity of the RUVBL1/2 complex. (F) HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated at 1 hpi with DMSO or increasing concentrations of CB-6644. Viral titres were quantified at 0 hpi and 6 hpi. Viral titres in CB-6644-treated cells at 6 hpi are shown as individual points with means connected by a line. Mean viral titres of untreated cells at 0 hpi (input) and of DMSO-treated cells at 6 hpi are represented by dashed lines. (G) Cell viability of uninfected CB-6644-treated cells measured in parallel of the infection, presented as a percentage of the DMSO control. (H) RUVBL1 and RUVBL2 co-immunoprecipitate with RV-A16 2C or 2BC in the absence of other viral components. HeLa-H1 cells were transfected with constructs encoding FLAG-RUVBL1, HA-RUVBL2, and Myc-tagged RV NSPs (2C or 2BC) or Myc-GFP. Myc-tagged proteins were immunoprecipitated from cell lysates. Cell lysates (input) and immunoprecipitated fractions (α-Myc IP) were analysed by western blotting for Myc, FLAG, and HA. (*) HA-RUVBL2 overlaps with IgG heavy chain. FLAG-RUVBL1 and HA-RUVBL2 with Myc-2C, N=4; FLAG-RUVBL1 with Myc-2BC, N=3; HA-RUVBL2 with Myc-2BC, N=2. For all graph panels (D-G), data from 3-4 independent experiments are shown as individual points coded by shape, according to experimental replicate, together with means (connected by lines in F-G). Non-graph panels (H) show representative images. See also Figure S1.
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Jackson Laboratory conditional knockout ruvbl1 mice
(A) Workflow for the identification of RV-A16 2C interactors and their functional validation. HeLa-H1 cells were infected in triplicate with RV-A16 (MOI 20) for 4.5 h, 6 h or were left uninfected. Protein-protein interactions were then cross-linked with formaldehyde and cell lysates were immunoprecipitated using RV-A16 2C-specific or control antisera (2C-IP or control-IP). Eluates were TMT-labelled, multiplexed and analysed by LC-MS/MS. Differences between infected and uninfected conditions were assessed using two-sample Student’s t -test. Cellular proteins significantly enriched in infected 2C-IP fractions were further investigated for their role in RV-A16 replication through siRNA screening. The two most significant non-cytotoxic hits from the siRNA screen, <t>RUVBL1</t> and RUVBL2, were further validated using a small molecule inhibitor. (B-C) Proteomics analysis of 2C-IPs. RV-A16 2C was immunoprecipitated from infected cells and the associated proteins were analysed as described in A. (B) Volcano plots showing in red the cellular proteins significantly enriched in 2C-IPs at 4.5 hpi (top) and 6 hpi (bottom), compared to uninfected conditions. (C) Corresponding heatmap, also showing infected control-IPs. (D-E) siRNA screen of the 2C-IP hits. HeLa-H1 cells were transfected with siRNA pools targeting the 2C-IPs hits or a non-targeting (NT) control siRNA pool. (D) At 72 h post-transfection, cells were infected with RV-A16 (MOI 20), and viral titres were quantified at 6 hpi. Data are presented as a percentage of the mean NT control (dashed grey line). (E) Viability of uninfected cells at 72 h post-transfection, for siRNA significantly reducing viral replication, presented as percentage of the mean NT control (dashed grey line). Statistical tests: one-way ANOVA with Dunnett’s post-hoc test, comparing to the NT control pool. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (F-G) Validation using CB-6644, a small molecule inhibitor of the ATPase activity of the RUVBL1/2 complex. (F) HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated at 1 hpi with DMSO or increasing concentrations of CB-6644. Viral titres were quantified at 0 hpi and 6 hpi. Viral titres in CB-6644-treated cells at 6 hpi are shown as individual points with means connected by a line. Mean viral titres of untreated cells at 0 hpi (input) and of DMSO-treated cells at 6 hpi are represented by dashed lines. (G) Cell viability of uninfected CB-6644-treated cells measured in parallel of the infection, presented as a percentage of the DMSO control. (H) RUVBL1 and RUVBL2 co-immunoprecipitate with RV-A16 2C or 2BC in the absence of other viral components. HeLa-H1 cells were transfected with constructs encoding FLAG-RUVBL1, HA-RUVBL2, and Myc-tagged RV NSPs (2C or 2BC) or Myc-GFP. Myc-tagged proteins were immunoprecipitated from cell lysates. Cell lysates (input) and immunoprecipitated fractions (α-Myc IP) were analysed by western blotting for Myc, FLAG, and HA. (*) HA-RUVBL2 overlaps with IgG heavy chain. FLAG-RUVBL1 and HA-RUVBL2 with Myc-2C, N=4; FLAG-RUVBL1 with Myc-2BC, N=3; HA-RUVBL2 with Myc-2BC, N=2. For all graph panels (D-G), data from 3-4 independent experiments are shown as individual points coded by shape, according to experimental replicate, together with means (connected by lines in F-G). Non-graph panels (H) show representative images. See also Figure S1.
Conditional Knockout Ruvbl1 Mice, supplied by Jackson Laboratory, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Cell Signaling Technology Inc anti ruvbl1
(A) Workflow for the identification of RV-A16 2C interactors and their functional validation. HeLa-H1 cells were infected in triplicate with RV-A16 (MOI 20) for 4.5 h, 6 h or were left uninfected. Protein-protein interactions were then cross-linked with formaldehyde and cell lysates were immunoprecipitated using RV-A16 2C-specific or control antisera (2C-IP or control-IP). Eluates were TMT-labelled, multiplexed and analysed by LC-MS/MS. Differences between infected and uninfected conditions were assessed using two-sample Student’s t -test. Cellular proteins significantly enriched in infected 2C-IP fractions were further investigated for their role in RV-A16 replication through siRNA screening. The two most significant non-cytotoxic hits from the siRNA screen, <t>RUVBL1</t> and RUVBL2, were further validated using a small molecule inhibitor. (B-C) Proteomics analysis of 2C-IPs. RV-A16 2C was immunoprecipitated from infected cells and the associated proteins were analysed as described in A. (B) Volcano plots showing in red the cellular proteins significantly enriched in 2C-IPs at 4.5 hpi (top) and 6 hpi (bottom), compared to uninfected conditions. (C) Corresponding heatmap, also showing infected control-IPs. (D-E) siRNA screen of the 2C-IP hits. HeLa-H1 cells were transfected with siRNA pools targeting the 2C-IPs hits or a non-targeting (NT) control siRNA pool. (D) At 72 h post-transfection, cells were infected with RV-A16 (MOI 20), and viral titres were quantified at 6 hpi. Data are presented as a percentage of the mean NT control (dashed grey line). (E) Viability of uninfected cells at 72 h post-transfection, for siRNA significantly reducing viral replication, presented as percentage of the mean NT control (dashed grey line). Statistical tests: one-way ANOVA with Dunnett’s post-hoc test, comparing to the NT control pool. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (F-G) Validation using CB-6644, a small molecule inhibitor of the ATPase activity of the RUVBL1/2 complex. (F) HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated at 1 hpi with DMSO or increasing concentrations of CB-6644. Viral titres were quantified at 0 hpi and 6 hpi. Viral titres in CB-6644-treated cells at 6 hpi are shown as individual points with means connected by a line. Mean viral titres of untreated cells at 0 hpi (input) and of DMSO-treated cells at 6 hpi are represented by dashed lines. (G) Cell viability of uninfected CB-6644-treated cells measured in parallel of the infection, presented as a percentage of the DMSO control. (H) RUVBL1 and RUVBL2 co-immunoprecipitate with RV-A16 2C or 2BC in the absence of other viral components. HeLa-H1 cells were transfected with constructs encoding FLAG-RUVBL1, HA-RUVBL2, and Myc-tagged RV NSPs (2C or 2BC) or Myc-GFP. Myc-tagged proteins were immunoprecipitated from cell lysates. Cell lysates (input) and immunoprecipitated fractions (α-Myc IP) were analysed by western blotting for Myc, FLAG, and HA. (*) HA-RUVBL2 overlaps with IgG heavy chain. FLAG-RUVBL1 and HA-RUVBL2 with Myc-2C, N=4; FLAG-RUVBL1 with Myc-2BC, N=3; HA-RUVBL2 with Myc-2BC, N=2. For all graph panels (D-G), data from 3-4 independent experiments are shown as individual points coded by shape, according to experimental replicate, together with means (connected by lines in F-G). Non-graph panels (H) show representative images. See also Figure S1.
Anti Ruvbl1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Cell Signaling Technology Inc anti pontin
(A) Workflow for the identification of RV-A16 2C interactors and their functional validation. HeLa-H1 cells were infected in triplicate with RV-A16 (MOI 20) for 4.5 h, 6 h or were left uninfected. Protein-protein interactions were then cross-linked with formaldehyde and cell lysates were immunoprecipitated using RV-A16 2C-specific or control antisera (2C-IP or control-IP). Eluates were TMT-labelled, multiplexed and analysed by LC-MS/MS. Differences between infected and uninfected conditions were assessed using two-sample Student’s t -test. Cellular proteins significantly enriched in infected 2C-IP fractions were further investigated for their role in RV-A16 replication through siRNA screening. The two most significant non-cytotoxic hits from the siRNA screen, <t>RUVBL1</t> and RUVBL2, were further validated using a small molecule inhibitor. (B-C) Proteomics analysis of 2C-IPs. RV-A16 2C was immunoprecipitated from infected cells and the associated proteins were analysed as described in A. (B) Volcano plots showing in red the cellular proteins significantly enriched in 2C-IPs at 4.5 hpi (top) and 6 hpi (bottom), compared to uninfected conditions. (C) Corresponding heatmap, also showing infected control-IPs. (D-E) siRNA screen of the 2C-IP hits. HeLa-H1 cells were transfected with siRNA pools targeting the 2C-IPs hits or a non-targeting (NT) control siRNA pool. (D) At 72 h post-transfection, cells were infected with RV-A16 (MOI 20), and viral titres were quantified at 6 hpi. Data are presented as a percentage of the mean NT control (dashed grey line). (E) Viability of uninfected cells at 72 h post-transfection, for siRNA significantly reducing viral replication, presented as percentage of the mean NT control (dashed grey line). Statistical tests: one-way ANOVA with Dunnett’s post-hoc test, comparing to the NT control pool. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (F-G) Validation using CB-6644, a small molecule inhibitor of the ATPase activity of the RUVBL1/2 complex. (F) HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated at 1 hpi with DMSO or increasing concentrations of CB-6644. Viral titres were quantified at 0 hpi and 6 hpi. Viral titres in CB-6644-treated cells at 6 hpi are shown as individual points with means connected by a line. Mean viral titres of untreated cells at 0 hpi (input) and of DMSO-treated cells at 6 hpi are represented by dashed lines. (G) Cell viability of uninfected CB-6644-treated cells measured in parallel of the infection, presented as a percentage of the DMSO control. (H) RUVBL1 and RUVBL2 co-immunoprecipitate with RV-A16 2C or 2BC in the absence of other viral components. HeLa-H1 cells were transfected with constructs encoding FLAG-RUVBL1, HA-RUVBL2, and Myc-tagged RV NSPs (2C or 2BC) or Myc-GFP. Myc-tagged proteins were immunoprecipitated from cell lysates. Cell lysates (input) and immunoprecipitated fractions (α-Myc IP) were analysed by western blotting for Myc, FLAG, and HA. (*) HA-RUVBL2 overlaps with IgG heavy chain. FLAG-RUVBL1 and HA-RUVBL2 with Myc-2C, N=4; FLAG-RUVBL1 with Myc-2BC, N=3; HA-RUVBL2 with Myc-2BC, N=2. For all graph panels (D-G), data from 3-4 independent experiments are shown as individual points coded by shape, according to experimental replicate, together with means (connected by lines in F-G). Non-graph panels (H) show representative images. See also Figure S1.
Anti Pontin, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Proteintech ruvbl1
(A) Workflow for the identification of RV-A16 2C interactors and their functional validation. HeLa-H1 cells were infected in triplicate with RV-A16 (MOI 20) for 4.5 h, 6 h or were left uninfected. Protein-protein interactions were then cross-linked with formaldehyde and cell lysates were immunoprecipitated using RV-A16 2C-specific or control antisera (2C-IP or control-IP). Eluates were TMT-labelled, multiplexed and analysed by LC-MS/MS. Differences between infected and uninfected conditions were assessed using two-sample Student’s t -test. Cellular proteins significantly enriched in infected 2C-IP fractions were further investigated for their role in RV-A16 replication through siRNA screening. The two most significant non-cytotoxic hits from the siRNA screen, <t>RUVBL1</t> and RUVBL2, were further validated using a small molecule inhibitor. (B-C) Proteomics analysis of 2C-IPs. RV-A16 2C was immunoprecipitated from infected cells and the associated proteins were analysed as described in A. (B) Volcano plots showing in red the cellular proteins significantly enriched in 2C-IPs at 4.5 hpi (top) and 6 hpi (bottom), compared to uninfected conditions. (C) Corresponding heatmap, also showing infected control-IPs. (D-E) siRNA screen of the 2C-IP hits. HeLa-H1 cells were transfected with siRNA pools targeting the 2C-IPs hits or a non-targeting (NT) control siRNA pool. (D) At 72 h post-transfection, cells were infected with RV-A16 (MOI 20), and viral titres were quantified at 6 hpi. Data are presented as a percentage of the mean NT control (dashed grey line). (E) Viability of uninfected cells at 72 h post-transfection, for siRNA significantly reducing viral replication, presented as percentage of the mean NT control (dashed grey line). Statistical tests: one-way ANOVA with Dunnett’s post-hoc test, comparing to the NT control pool. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (F-G) Validation using CB-6644, a small molecule inhibitor of the ATPase activity of the RUVBL1/2 complex. (F) HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated at 1 hpi with DMSO or increasing concentrations of CB-6644. Viral titres were quantified at 0 hpi and 6 hpi. Viral titres in CB-6644-treated cells at 6 hpi are shown as individual points with means connected by a line. Mean viral titres of untreated cells at 0 hpi (input) and of DMSO-treated cells at 6 hpi are represented by dashed lines. (G) Cell viability of uninfected CB-6644-treated cells measured in parallel of the infection, presented as a percentage of the DMSO control. (H) RUVBL1 and RUVBL2 co-immunoprecipitate with RV-A16 2C or 2BC in the absence of other viral components. HeLa-H1 cells were transfected with constructs encoding FLAG-RUVBL1, HA-RUVBL2, and Myc-tagged RV NSPs (2C or 2BC) or Myc-GFP. Myc-tagged proteins were immunoprecipitated from cell lysates. Cell lysates (input) and immunoprecipitated fractions (α-Myc IP) were analysed by western blotting for Myc, FLAG, and HA. (*) HA-RUVBL2 overlaps with IgG heavy chain. FLAG-RUVBL1 and HA-RUVBL2 with Myc-2C, N=4; FLAG-RUVBL1 with Myc-2BC, N=3; HA-RUVBL2 with Myc-2BC, N=2. For all graph panels (D-G), data from 3-4 independent experiments are shown as individual points coded by shape, according to experimental replicate, together with means (connected by lines in F-G). Non-graph panels (H) show representative images. See also Figure S1.
Ruvbl1, supplied by Proteintech, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Proteintech anti ruvbl1
(A) Workflow for the identification of RV-A16 2C interactors and their functional validation. HeLa-H1 cells were infected in triplicate with RV-A16 (MOI 20) for 4.5 h, 6 h or were left uninfected. Protein-protein interactions were then cross-linked with formaldehyde and cell lysates were immunoprecipitated using RV-A16 2C-specific or control antisera (2C-IP or control-IP). Eluates were TMT-labelled, multiplexed and analysed by LC-MS/MS. Differences between infected and uninfected conditions were assessed using two-sample Student’s t -test. Cellular proteins significantly enriched in infected 2C-IP fractions were further investigated for their role in RV-A16 replication through siRNA screening. The two most significant non-cytotoxic hits from the siRNA screen, <t>RUVBL1</t> and RUVBL2, were further validated using a small molecule inhibitor. (B-C) Proteomics analysis of 2C-IPs. RV-A16 2C was immunoprecipitated from infected cells and the associated proteins were analysed as described in A. (B) Volcano plots showing in red the cellular proteins significantly enriched in 2C-IPs at 4.5 hpi (top) and 6 hpi (bottom), compared to uninfected conditions. (C) Corresponding heatmap, also showing infected control-IPs. (D-E) siRNA screen of the 2C-IP hits. HeLa-H1 cells were transfected with siRNA pools targeting the 2C-IPs hits or a non-targeting (NT) control siRNA pool. (D) At 72 h post-transfection, cells were infected with RV-A16 (MOI 20), and viral titres were quantified at 6 hpi. Data are presented as a percentage of the mean NT control (dashed grey line). (E) Viability of uninfected cells at 72 h post-transfection, for siRNA significantly reducing viral replication, presented as percentage of the mean NT control (dashed grey line). Statistical tests: one-way ANOVA with Dunnett’s post-hoc test, comparing to the NT control pool. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (F-G) Validation using CB-6644, a small molecule inhibitor of the ATPase activity of the RUVBL1/2 complex. (F) HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated at 1 hpi with DMSO or increasing concentrations of CB-6644. Viral titres were quantified at 0 hpi and 6 hpi. Viral titres in CB-6644-treated cells at 6 hpi are shown as individual points with means connected by a line. Mean viral titres of untreated cells at 0 hpi (input) and of DMSO-treated cells at 6 hpi are represented by dashed lines. (G) Cell viability of uninfected CB-6644-treated cells measured in parallel of the infection, presented as a percentage of the DMSO control. (H) RUVBL1 and RUVBL2 co-immunoprecipitate with RV-A16 2C or 2BC in the absence of other viral components. HeLa-H1 cells were transfected with constructs encoding FLAG-RUVBL1, HA-RUVBL2, and Myc-tagged RV NSPs (2C or 2BC) or Myc-GFP. Myc-tagged proteins were immunoprecipitated from cell lysates. Cell lysates (input) and immunoprecipitated fractions (α-Myc IP) were analysed by western blotting for Myc, FLAG, and HA. (*) HA-RUVBL2 overlaps with IgG heavy chain. FLAG-RUVBL1 and HA-RUVBL2 with Myc-2C, N=4; FLAG-RUVBL1 with Myc-2BC, N=3; HA-RUVBL2 with Myc-2BC, N=2. For all graph panels (D-G), data from 3-4 independent experiments are shown as individual points coded by shape, according to experimental replicate, together with means (connected by lines in F-G). Non-graph panels (H) show representative images. See also Figure S1.
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(A) Workflow for the identification of RV-A16 2C interactors and their functional validation. HeLa-H1 cells were infected in triplicate with RV-A16 (MOI 20) for 4.5 h, 6 h or were left uninfected. Protein-protein interactions were then cross-linked with formaldehyde and cell lysates were immunoprecipitated using RV-A16 2C-specific or control antisera (2C-IP or control-IP). Eluates were TMT-labelled, multiplexed and analysed by LC-MS/MS. Differences between infected and uninfected conditions were assessed using two-sample Student’s t -test. Cellular proteins significantly enriched in infected 2C-IP fractions were further investigated for their role in RV-A16 replication through siRNA screening. The two most significant non-cytotoxic hits from the siRNA screen, <t>RUVBL1</t> and RUVBL2, were further validated using a small molecule inhibitor. (B-C) Proteomics analysis of 2C-IPs. RV-A16 2C was immunoprecipitated from infected cells and the associated proteins were analysed as described in A. (B) Volcano plots showing in red the cellular proteins significantly enriched in 2C-IPs at 4.5 hpi (top) and 6 hpi (bottom), compared to uninfected conditions. (C) Corresponding heatmap, also showing infected control-IPs. (D-E) siRNA screen of the 2C-IP hits. HeLa-H1 cells were transfected with siRNA pools targeting the 2C-IPs hits or a non-targeting (NT) control siRNA pool. (D) At 72 h post-transfection, cells were infected with RV-A16 (MOI 20), and viral titres were quantified at 6 hpi. Data are presented as a percentage of the mean NT control (dashed grey line). (E) Viability of uninfected cells at 72 h post-transfection, for siRNA significantly reducing viral replication, presented as percentage of the mean NT control (dashed grey line). Statistical tests: one-way ANOVA with Dunnett’s post-hoc test, comparing to the NT control pool. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (F-G) Validation using CB-6644, a small molecule inhibitor of the ATPase activity of the RUVBL1/2 complex. (F) HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated at 1 hpi with DMSO or increasing concentrations of CB-6644. Viral titres were quantified at 0 hpi and 6 hpi. Viral titres in CB-6644-treated cells at 6 hpi are shown as individual points with means connected by a line. Mean viral titres of untreated cells at 0 hpi (input) and of DMSO-treated cells at 6 hpi are represented by dashed lines. (G) Cell viability of uninfected CB-6644-treated cells measured in parallel of the infection, presented as a percentage of the DMSO control. (H) RUVBL1 and RUVBL2 co-immunoprecipitate with RV-A16 2C or 2BC in the absence of other viral components. HeLa-H1 cells were transfected with constructs encoding FLAG-RUVBL1, HA-RUVBL2, and Myc-tagged RV NSPs (2C or 2BC) or Myc-GFP. Myc-tagged proteins were immunoprecipitated from cell lysates. Cell lysates (input) and immunoprecipitated fractions (α-Myc IP) were analysed by western blotting for Myc, FLAG, and HA. (*) HA-RUVBL2 overlaps with IgG heavy chain. FLAG-RUVBL1 and HA-RUVBL2 with Myc-2C, N=4; FLAG-RUVBL1 with Myc-2BC, N=3; HA-RUVBL2 with Myc-2BC, N=2. For all graph panels (D-G), data from 3-4 independent experiments are shown as individual points coded by shape, according to experimental replicate, together with means (connected by lines in F-G). Non-graph panels (H) show representative images. See also Figure S1.
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Image Search Results


(A) Workflow for the identification of RV-A16 2C interactors and their functional validation. HeLa-H1 cells were infected in triplicate with RV-A16 (MOI 20) for 4.5 h, 6 h or were left uninfected. Protein-protein interactions were then cross-linked with formaldehyde and cell lysates were immunoprecipitated using RV-A16 2C-specific or control antisera (2C-IP or control-IP). Eluates were TMT-labelled, multiplexed and analysed by LC-MS/MS. Differences between infected and uninfected conditions were assessed using two-sample Student’s t -test. Cellular proteins significantly enriched in infected 2C-IP fractions were further investigated for their role in RV-A16 replication through siRNA screening. The two most significant non-cytotoxic hits from the siRNA screen, RUVBL1 and RUVBL2, were further validated using a small molecule inhibitor. (B-C) Proteomics analysis of 2C-IPs. RV-A16 2C was immunoprecipitated from infected cells and the associated proteins were analysed as described in A. (B) Volcano plots showing in red the cellular proteins significantly enriched in 2C-IPs at 4.5 hpi (top) and 6 hpi (bottom), compared to uninfected conditions. (C) Corresponding heatmap, also showing infected control-IPs. (D-E) siRNA screen of the 2C-IP hits. HeLa-H1 cells were transfected with siRNA pools targeting the 2C-IPs hits or a non-targeting (NT) control siRNA pool. (D) At 72 h post-transfection, cells were infected with RV-A16 (MOI 20), and viral titres were quantified at 6 hpi. Data are presented as a percentage of the mean NT control (dashed grey line). (E) Viability of uninfected cells at 72 h post-transfection, for siRNA significantly reducing viral replication, presented as percentage of the mean NT control (dashed grey line). Statistical tests: one-way ANOVA with Dunnett’s post-hoc test, comparing to the NT control pool. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (F-G) Validation using CB-6644, a small molecule inhibitor of the ATPase activity of the RUVBL1/2 complex. (F) HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated at 1 hpi with DMSO or increasing concentrations of CB-6644. Viral titres were quantified at 0 hpi and 6 hpi. Viral titres in CB-6644-treated cells at 6 hpi are shown as individual points with means connected by a line. Mean viral titres of untreated cells at 0 hpi (input) and of DMSO-treated cells at 6 hpi are represented by dashed lines. (G) Cell viability of uninfected CB-6644-treated cells measured in parallel of the infection, presented as a percentage of the DMSO control. (H) RUVBL1 and RUVBL2 co-immunoprecipitate with RV-A16 2C or 2BC in the absence of other viral components. HeLa-H1 cells were transfected with constructs encoding FLAG-RUVBL1, HA-RUVBL2, and Myc-tagged RV NSPs (2C or 2BC) or Myc-GFP. Myc-tagged proteins were immunoprecipitated from cell lysates. Cell lysates (input) and immunoprecipitated fractions (α-Myc IP) were analysed by western blotting for Myc, FLAG, and HA. (*) HA-RUVBL2 overlaps with IgG heavy chain. FLAG-RUVBL1 and HA-RUVBL2 with Myc-2C, N=4; FLAG-RUVBL1 with Myc-2BC, N=3; HA-RUVBL2 with Myc-2BC, N=2. For all graph panels (D-G), data from 3-4 independent experiments are shown as individual points coded by shape, according to experimental replicate, together with means (connected by lines in F-G). Non-graph panels (H) show representative images. See also Figure S1.

Journal: bioRxiv

Article Title: A host ATPase essential for rhinovirus replication is an antiviral target with a high barrier to resistance

doi: 10.64898/2026.05.13.723454

Figure Lengend Snippet: (A) Workflow for the identification of RV-A16 2C interactors and their functional validation. HeLa-H1 cells were infected in triplicate with RV-A16 (MOI 20) for 4.5 h, 6 h or were left uninfected. Protein-protein interactions were then cross-linked with formaldehyde and cell lysates were immunoprecipitated using RV-A16 2C-specific or control antisera (2C-IP or control-IP). Eluates were TMT-labelled, multiplexed and analysed by LC-MS/MS. Differences between infected and uninfected conditions were assessed using two-sample Student’s t -test. Cellular proteins significantly enriched in infected 2C-IP fractions were further investigated for their role in RV-A16 replication through siRNA screening. The two most significant non-cytotoxic hits from the siRNA screen, RUVBL1 and RUVBL2, were further validated using a small molecule inhibitor. (B-C) Proteomics analysis of 2C-IPs. RV-A16 2C was immunoprecipitated from infected cells and the associated proteins were analysed as described in A. (B) Volcano plots showing in red the cellular proteins significantly enriched in 2C-IPs at 4.5 hpi (top) and 6 hpi (bottom), compared to uninfected conditions. (C) Corresponding heatmap, also showing infected control-IPs. (D-E) siRNA screen of the 2C-IP hits. HeLa-H1 cells were transfected with siRNA pools targeting the 2C-IPs hits or a non-targeting (NT) control siRNA pool. (D) At 72 h post-transfection, cells were infected with RV-A16 (MOI 20), and viral titres were quantified at 6 hpi. Data are presented as a percentage of the mean NT control (dashed grey line). (E) Viability of uninfected cells at 72 h post-transfection, for siRNA significantly reducing viral replication, presented as percentage of the mean NT control (dashed grey line). Statistical tests: one-way ANOVA with Dunnett’s post-hoc test, comparing to the NT control pool. **, P < 0.01; ***, P < 0.001; ****, P < 0.0001. (F-G) Validation using CB-6644, a small molecule inhibitor of the ATPase activity of the RUVBL1/2 complex. (F) HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated at 1 hpi with DMSO or increasing concentrations of CB-6644. Viral titres were quantified at 0 hpi and 6 hpi. Viral titres in CB-6644-treated cells at 6 hpi are shown as individual points with means connected by a line. Mean viral titres of untreated cells at 0 hpi (input) and of DMSO-treated cells at 6 hpi are represented by dashed lines. (G) Cell viability of uninfected CB-6644-treated cells measured in parallel of the infection, presented as a percentage of the DMSO control. (H) RUVBL1 and RUVBL2 co-immunoprecipitate with RV-A16 2C or 2BC in the absence of other viral components. HeLa-H1 cells were transfected with constructs encoding FLAG-RUVBL1, HA-RUVBL2, and Myc-tagged RV NSPs (2C or 2BC) or Myc-GFP. Myc-tagged proteins were immunoprecipitated from cell lysates. Cell lysates (input) and immunoprecipitated fractions (α-Myc IP) were analysed by western blotting for Myc, FLAG, and HA. (*) HA-RUVBL2 overlaps with IgG heavy chain. FLAG-RUVBL1 and HA-RUVBL2 with Myc-2C, N=4; FLAG-RUVBL1 with Myc-2BC, N=3; HA-RUVBL2 with Myc-2BC, N=2. For all graph panels (D-G), data from 3-4 independent experiments are shown as individual points coded by shape, according to experimental replicate, together with means (connected by lines in F-G). Non-graph panels (H) show representative images. See also Figure S1.

Article Snippet: Single siRNA targeting RUVBL1 and firefly luciferase were purchased from Eurofins.

Techniques: Functional Assay, Biomarker Discovery, Infection, Protein-Protein interactions, Immunoprecipitation, Control, Liquid Chromatography with Mass Spectroscopy, Transfection, Activity Assay, Construct, Western Blot

(A–C) CB-6644 inhibits RV RNA replication and NSP production. HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated at 1 hpi with DMSO or 500 nM CB-6644. (A) Viral RNA was quantified by RT-qPCR at 0 hpi and 6 hpi. (B) At 6 hpi, lysates were analysed by western blotting for RV-A16 3C, RUVBL1, and lamin-B1. (C) 3C signal was quantified and normalised to lamin-B1. (D) Time-of-addition assay. HeLa-H1 cells were infected as above and treated with DMSO or 500 nM CB-6644 immediately after virus adsorption (0 hpi) or at the indicated times post-infection. Viral titres were quantified at 6 hpi. (E–J) siRNA knockdown of RUVBL1 inhibits RV RNA replication and NSP production. HeLa-H1 cells were transfected with siRNA targeting RUVBL1 or firefly luciferase for 72 h and then infected with RV-A16 (MOI 20). (E) Viral RNA was quantified by RT-qPCR at 0 hpi and 6 hpi. (F) At 6 hpi, lysates were analysed by western blotting for RV-A16 3C, RUVBL1, and lamin-B1. (G–H) Quantification of 3C and RUVBL1 signal from F, normalised to lamin-B1. (I) Immunofluorescence staining for RV-A16 2C (red) at 6 hpi; nuclei were stained with DAPI (blue). (J) Quantification of 2C-positive cells from (I). (K–L) RUVBL1/2 is required after RV entry. (K) HeLa-H1 cells were transfected with RV-A16, RV-A1a, or RV-B14 RNA in the presence of DMSO or 500 nM CB-6644. Viral titres were quantified at 14 h post-transfection. (L) Cell viability assessed in parallel of K in untransfected cells treated for 14 h with DMSO or 500 nM CB-6644. (M) RUVBL1/2 is not required for IRES-dependent translation. HeLa-H1 cells were transfected with a luciferase reporter RNA under RV-A16 IRES-dependent translational control, in the presence of DMSO, 500 nM CB-6644, or cycloheximide (CHX). Luciferase activity was measured at the indicated times. Values were t=0-subtracted and normalised to the DMSO 24 h post-transfection value within each experiment. The 0 h baseline is shown as a dashed grey line. (N-O) RUVBL1/2 is not required for RV-A16 polyprotein cleavage. (N) Myc-GFP-2BC3ABCD construct used for polyprotein processing assays, with expression under the control of a CMV promoter. (O) HeLa-H1 cells were transfected or not with the Myc-GFP-2BC3ABCD plasmid for 21 h, in the presence of DMSO or 500 nM CB-6644. In parallel, HeLa-H1 cells were infected or not with RV-A16 for 8 h. Lysates were then analysed by western blotting for Myc-GFP and RV-A16 2C, 3A and 3C. (P) CB-6644 inhibits negative-strand RNA synthesis. HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated with DMSO or 500 nM CB-6644 at 1 hpi. Negative-strand RNA was quantified at the indicated times by RT-qPCR, normalised to 0 hpi. For all graph panels (A, C-E, G, H, J-M, P), data from 3-4 independent experiments are shown as individual points, coded by shape according to experimental replicate, with means (connected by lines in M and P). Non-graph panels (B, F, I, O) show representative images from 3 independent experiments. Statistical tests: two-tailed paired t-test (A, C, E, G, H, J-L), one-way ANOVA with Dunnett’s post-hoc test (D), two-way ANOVA, comparing drug treatments to the DMSO control at each time point (M, P). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant. See also Figure S2 and S3.

Journal: bioRxiv

Article Title: A host ATPase essential for rhinovirus replication is an antiviral target with a high barrier to resistance

doi: 10.64898/2026.05.13.723454

Figure Lengend Snippet: (A–C) CB-6644 inhibits RV RNA replication and NSP production. HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated at 1 hpi with DMSO or 500 nM CB-6644. (A) Viral RNA was quantified by RT-qPCR at 0 hpi and 6 hpi. (B) At 6 hpi, lysates were analysed by western blotting for RV-A16 3C, RUVBL1, and lamin-B1. (C) 3C signal was quantified and normalised to lamin-B1. (D) Time-of-addition assay. HeLa-H1 cells were infected as above and treated with DMSO or 500 nM CB-6644 immediately after virus adsorption (0 hpi) or at the indicated times post-infection. Viral titres were quantified at 6 hpi. (E–J) siRNA knockdown of RUVBL1 inhibits RV RNA replication and NSP production. HeLa-H1 cells were transfected with siRNA targeting RUVBL1 or firefly luciferase for 72 h and then infected with RV-A16 (MOI 20). (E) Viral RNA was quantified by RT-qPCR at 0 hpi and 6 hpi. (F) At 6 hpi, lysates were analysed by western blotting for RV-A16 3C, RUVBL1, and lamin-B1. (G–H) Quantification of 3C and RUVBL1 signal from F, normalised to lamin-B1. (I) Immunofluorescence staining for RV-A16 2C (red) at 6 hpi; nuclei were stained with DAPI (blue). (J) Quantification of 2C-positive cells from (I). (K–L) RUVBL1/2 is required after RV entry. (K) HeLa-H1 cells were transfected with RV-A16, RV-A1a, or RV-B14 RNA in the presence of DMSO or 500 nM CB-6644. Viral titres were quantified at 14 h post-transfection. (L) Cell viability assessed in parallel of K in untransfected cells treated for 14 h with DMSO or 500 nM CB-6644. (M) RUVBL1/2 is not required for IRES-dependent translation. HeLa-H1 cells were transfected with a luciferase reporter RNA under RV-A16 IRES-dependent translational control, in the presence of DMSO, 500 nM CB-6644, or cycloheximide (CHX). Luciferase activity was measured at the indicated times. Values were t=0-subtracted and normalised to the DMSO 24 h post-transfection value within each experiment. The 0 h baseline is shown as a dashed grey line. (N-O) RUVBL1/2 is not required for RV-A16 polyprotein cleavage. (N) Myc-GFP-2BC3ABCD construct used for polyprotein processing assays, with expression under the control of a CMV promoter. (O) HeLa-H1 cells were transfected or not with the Myc-GFP-2BC3ABCD plasmid for 21 h, in the presence of DMSO or 500 nM CB-6644. In parallel, HeLa-H1 cells were infected or not with RV-A16 for 8 h. Lysates were then analysed by western blotting for Myc-GFP and RV-A16 2C, 3A and 3C. (P) CB-6644 inhibits negative-strand RNA synthesis. HeLa-H1 cells were infected with RV-A16 (MOI 20) and treated with DMSO or 500 nM CB-6644 at 1 hpi. Negative-strand RNA was quantified at the indicated times by RT-qPCR, normalised to 0 hpi. For all graph panels (A, C-E, G, H, J-M, P), data from 3-4 independent experiments are shown as individual points, coded by shape according to experimental replicate, with means (connected by lines in M and P). Non-graph panels (B, F, I, O) show representative images from 3 independent experiments. Statistical tests: two-tailed paired t-test (A, C, E, G, H, J-L), one-way ANOVA with Dunnett’s post-hoc test (D), two-way ANOVA, comparing drug treatments to the DMSO control at each time point (M, P). *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant. See also Figure S2 and S3.

Article Snippet: Single siRNA targeting RUVBL1 and firefly luciferase were purchased from Eurofins.

Techniques: Infection, Quantitative RT-PCR, Western Blot, Virus, Adsorption, Knockdown, Transfection, Luciferase, Immunofluorescence, Staining, Control, Activity Assay, Construct, Expressing, Plasmid Preparation, Two Tailed Test