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ebola minigenome assay plasmids  (Addgene inc)
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(A) Schematic overview of the <t>minigenome</t> system, tested conditions and workflow. HEK293T cells were transfected with EBOV RNP plasmids encoding L, NP, VP35, VP30 proteins and a eGFP-reporter minigenome (MG). Following transcription by host RNA polymerase II, viral RNA is replicated and transcribed by the RNP complex, with eGFP expression serving as a readout of replication/transcription activity. Four conditions were analyzed: L (functional ribonuclear complex), mutL (inactive polymerase L with N743A mutation in catalytic domain), −VP30, and NC (MG only). Cells were collected at 6 and 24 hrs post-transfection, followed by proteome and phosphoproteome analysis by LC-MS/MS. (B) Protein rank plot of L condition shows the EBOV RNP proteins (pink) within the proteome rank. (C,D) Volcano plots represent Log2-transformed abundance fold change (FC) of L condition versus NC 24 H post-transfection for protein groups (C) and phosphosites (D). Mint green represents the increase in abundance of host proteins in L condition (FC > 1, p-value <0.05). Brown represents the decrease in abundance of host proteins in L condition (FC < -1, p-value <0.05). Pink highlights EBOV proteins or EBOV protein phosphosites. Overall, 7447 protein groups and 10126 phosphosites could be quantified. (E) Regulated proteins and phosphosites that are either previously associated with EBOV infection or key components of enriched WikiPathway “EBOV infection in host” (p-value<0.05). The abundances were standardized using Z-scoring.
Ebola Minigenome Assay Plasmids, supplied by Addgene inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A) Schematic overview of the <t>minigenome</t> system, tested conditions and workflow. HEK293T cells were transfected with EBOV RNP plasmids encoding L, NP, VP35, VP30 proteins and a eGFP-reporter minigenome (MG). Following transcription by host RNA polymerase II, viral RNA is replicated and transcribed by the RNP complex, with eGFP expression serving as a readout of replication/transcription activity. Four conditions were analyzed: L (functional ribonuclear complex), mutL (inactive polymerase L with N743A mutation in catalytic domain), −VP30, and NC (MG only). Cells were collected at 6 and 24 hrs post-transfection, followed by proteome and phosphoproteome analysis by LC-MS/MS. (B) Protein rank plot of L condition shows the EBOV RNP proteins (pink) within the proteome rank. (C,D) Volcano plots represent Log2-transformed abundance fold change (FC) of L condition versus NC 24 H post-transfection for protein groups (C) and phosphosites (D). Mint green represents the increase in abundance of host proteins in L condition (FC > 1, p-value <0.05). Brown represents the decrease in abundance of host proteins in L condition (FC < -1, p-value <0.05). Pink highlights EBOV proteins or EBOV protein phosphosites. Overall, 7447 protein groups and 10126 phosphosites could be quantified. (E) Regulated proteins and phosphosites that are either previously associated with EBOV infection or key components of enriched WikiPathway “EBOV infection in host” (p-value<0.05). The abundances were standardized using Z-scoring.
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catalogue number plasmid 156438  (Addgene inc)
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(A) Schematic overview of the <t>minigenome</t> system, tested conditions and workflow. HEK293T cells were transfected with EBOV RNP plasmids encoding L, NP, VP35, VP30 proteins and a eGFP-reporter minigenome (MG). Following transcription by host RNA polymerase II, viral RNA is replicated and transcribed by the RNP complex, with eGFP expression serving as a readout of replication/transcription activity. Four conditions were analyzed: L (functional ribonuclear complex), mutL (inactive polymerase L with N743A mutation in catalytic domain), −VP30, and NC (MG only). Cells were collected at 6 and 24 hrs post-transfection, followed by proteome and phosphoproteome analysis by LC-MS/MS. (B) Protein rank plot of L condition shows the EBOV RNP proteins (pink) within the proteome rank. (C,D) Volcano plots represent Log2-transformed abundance fold change (FC) of L condition versus NC 24 H post-transfection for protein groups (C) and phosphosites (D). Mint green represents the increase in abundance of host proteins in L condition (FC > 1, p-value <0.05). Brown represents the decrease in abundance of host proteins in L condition (FC < -1, p-value <0.05). Pink highlights EBOV proteins or EBOV protein phosphosites. Overall, 7447 protein groups and 10126 phosphosites could be quantified. (E) Regulated proteins and phosphosites that are either previously associated with EBOV infection or key components of enriched WikiPathway “EBOV infection in host” (p-value<0.05). The abundances were standardized using Z-scoring.
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(A) Schematic overview of the <t>minigenome</t> system, tested conditions and workflow. HEK293T cells were transfected with EBOV RNP plasmids encoding L, NP, VP35, VP30 proteins and a eGFP-reporter minigenome (MG). Following transcription by host RNA polymerase II, viral RNA is replicated and transcribed by the RNP complex, with eGFP expression serving as a readout of replication/transcription activity. Four conditions were analyzed: L (functional ribonuclear complex), mutL (inactive polymerase L with N743A mutation in catalytic domain), −VP30, and NC (MG only). Cells were collected at 6 and 24 hrs post-transfection, followed by proteome and phosphoproteome analysis by LC-MS/MS. (B) Protein rank plot of L condition shows the EBOV RNP proteins (pink) within the proteome rank. (C,D) Volcano plots represent Log2-transformed abundance fold change (FC) of L condition versus NC 24 H post-transfection for protein groups (C) and phosphosites (D). Mint green represents the increase in abundance of host proteins in L condition (FC > 1, p-value <0.05). Brown represents the decrease in abundance of host proteins in L condition (FC < -1, p-value <0.05). Pink highlights EBOV proteins or EBOV protein phosphosites. Overall, 7447 protein groups and 10126 phosphosites could be quantified. (E) Regulated proteins and phosphosites that are either previously associated with EBOV infection or key components of enriched WikiPathway “EBOV infection in host” (p-value<0.05). The abundances were standardized using Z-scoring.
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(A) Schematic overview of the minigenome system, tested conditions and workflow. HEK293T cells were transfected with EBOV RNP plasmids encoding L, NP, VP35, VP30 proteins and a eGFP-reporter minigenome (MG). Following transcription by host RNA polymerase II, viral RNA is replicated and transcribed by the RNP complex, with eGFP expression serving as a readout of replication/transcription activity. Four conditions were analyzed: L (functional ribonuclear complex), mutL (inactive polymerase L with N743A mutation in catalytic domain), −VP30, and NC (MG only). Cells were collected at 6 and 24 hrs post-transfection, followed by proteome and phosphoproteome analysis by LC-MS/MS. (B) Protein rank plot of L condition shows the EBOV RNP proteins (pink) within the proteome rank. (C,D) Volcano plots represent Log2-transformed abundance fold change (FC) of L condition versus NC 24 H post-transfection for protein groups (C) and phosphosites (D). Mint green represents the increase in abundance of host proteins in L condition (FC > 1, p-value <0.05). Brown represents the decrease in abundance of host proteins in L condition (FC < -1, p-value <0.05). Pink highlights EBOV proteins or EBOV protein phosphosites. Overall, 7447 protein groups and 10126 phosphosites could be quantified. (E) Regulated proteins and phosphosites that are either previously associated with EBOV infection or key components of enriched WikiPathway “EBOV infection in host” (p-value<0.05). The abundances were standardized using Z-scoring.

Journal: bioRxiv

Article Title: Host cell remodeling via cyclin dependent kinases drives Ebola virus replication and transcription

doi: 10.64898/2026.03.25.714206

Figure Lengend Snippet: (A) Schematic overview of the minigenome system, tested conditions and workflow. HEK293T cells were transfected with EBOV RNP plasmids encoding L, NP, VP35, VP30 proteins and a eGFP-reporter minigenome (MG). Following transcription by host RNA polymerase II, viral RNA is replicated and transcribed by the RNP complex, with eGFP expression serving as a readout of replication/transcription activity. Four conditions were analyzed: L (functional ribonuclear complex), mutL (inactive polymerase L with N743A mutation in catalytic domain), −VP30, and NC (MG only). Cells were collected at 6 and 24 hrs post-transfection, followed by proteome and phosphoproteome analysis by LC-MS/MS. (B) Protein rank plot of L condition shows the EBOV RNP proteins (pink) within the proteome rank. (C,D) Volcano plots represent Log2-transformed abundance fold change (FC) of L condition versus NC 24 H post-transfection for protein groups (C) and phosphosites (D). Mint green represents the increase in abundance of host proteins in L condition (FC > 1, p-value <0.05). Brown represents the decrease in abundance of host proteins in L condition (FC < -1, p-value <0.05). Pink highlights EBOV proteins or EBOV protein phosphosites. Overall, 7447 protein groups and 10126 phosphosites could be quantified. (E) Regulated proteins and phosphosites that are either previously associated with EBOV infection or key components of enriched WikiPathway “EBOV infection in host” (p-value<0.05). The abundances were standardized using Z-scoring.

Article Snippet: The Ebola minigenome assay plasmids were purchased from Addgene: pCAGGS_3E5E_eGFP (ID: 103054); pCAGGS_L_EBOV (ID: 103052); pCAGGS_VP30_EBOV (ID: 103051); pCAGGS_VP35_EBOV (ID: 103050); pCAGGS_NP_EBOV (ID: 103049) ( ).

Techniques: Transfection, Expressing, Activity Assay, Functional Assay, Mutagenesis, Liquid Chromatography with Mass Spectroscopy, Transformation Assay, Infection

(A) Cell viability of cells treated with CDK inhibitors across a range of concentrations, relative to DMSO control. (B) Representative images of the HEK293T cell cultures treated with CDK2 (K03861; A674563), CDK4/6 (Palbociclib), CDK1 (Ro-3306) and panCDK (Flavopiridol) inhibitors or DMSO 1 H before or 4 H after the minigenome system transfection for 18 hours. The GFP signal represents cells with an active EBOV replication/transcription complex. Kinase inhibitor concentrations used for treatment are indicated in the lower right corner (0.5, 2.5, and 10 µM). Scale bars, 300 µm.

Journal: bioRxiv

Article Title: Host cell remodeling via cyclin dependent kinases drives Ebola virus replication and transcription

doi: 10.64898/2026.03.25.714206

Figure Lengend Snippet: (A) Cell viability of cells treated with CDK inhibitors across a range of concentrations, relative to DMSO control. (B) Representative images of the HEK293T cell cultures treated with CDK2 (K03861; A674563), CDK4/6 (Palbociclib), CDK1 (Ro-3306) and panCDK (Flavopiridol) inhibitors or DMSO 1 H before or 4 H after the minigenome system transfection for 18 hours. The GFP signal represents cells with an active EBOV replication/transcription complex. Kinase inhibitor concentrations used for treatment are indicated in the lower right corner (0.5, 2.5, and 10 µM). Scale bars, 300 µm.

Article Snippet: The Ebola minigenome assay plasmids were purchased from Addgene: pCAGGS_3E5E_eGFP (ID: 103054); pCAGGS_L_EBOV (ID: 103052); pCAGGS_VP30_EBOV (ID: 103051); pCAGGS_VP35_EBOV (ID: 103050); pCAGGS_NP_EBOV (ID: 103049) ( ).

Techniques: Control, Transfection