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poly i c  (InvivoGen)


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    InvivoGen poly i c
    ( A ) Schematic type I IFN pathway representation. Created in BioRender. Paeschke, K. (2026) https://BioRender.com/5jo0emg . ( B ) Representative confocal images (left) and quantification (right) of G3BP1-positive SGs (green) and DAPI (blue). The Mann-Whitney U test compared N = 683 WT with N = 1060 DHX36 KO cells; each point is the image mean of multiple cells ± SD. ( C and D ) Western blot quantification of phosphor-PKR and phosphor-eIF2α levels in WT and DHX36 KO cells. Unpaired two-tailed t test on N = 5 independent experiments ± SD. Representative blots are shown in fig. S1 [(A) and (B)]. ( E ) Evaluation of G3BP1 foci/cells by IF staining of the indicated cell lines. The Mann-Whitney U test compared N = 379 (WT) with N = 505 (DHX36 KO), N = 420 (PKR KO), and N = 358 (DHX36/PKR KO); each point is the image mean of multiple cells ± SD. Analysis corresponds to representative images in fig. S1E. ( F ) Western blot quantification of the phosphor-eIF2α levels in the indicated cell lines. Unpaired two-tailed t test, N ≥ 5 independent experiments ± SD. ( G ) Evaluation of CFAs of WT and DHX36 KO cells. Representative CFA images can be found in fig. S1F. ( H and I ) Cell confluency measurements of WT and DHX36 KO cells. Data from N = 6 independent experiments ± SE. ( J ) Evaluation of G3BP1-positive cells upon <t>poly(I:C)</t> treatment for the indicated periods by IF. The Mann-Whitney U test compared N ≥ 838 WT cells with N ≥ 1050 DHX36 KO cells ± SD for each condition. Representative confocal microscopy images are shown in fig. S1J. ( K ) Same as in (C) but with poly(I:C)-treated cells; Data from N = 3 independent experiments. Representative Western blot is shown in fig. S1K. ( L ) Same as in (F) but with poly(I:C)-treated cells.
    Poly I C, supplied by InvivoGen, used in various techniques. Bioz Stars score: 98/100, based on 4869 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/i+c+pic/pmc13220864-298-5-11?v=InvivoGen
    Average 98 stars, based on 4869 article reviews
    poly i c - by Bioz Stars, 2026-07
    98/100 stars

    Images

    1) Product Images from "DHX36 is a regulatory switch in the interferon-mediated antiviral response"

    Article Title: DHX36 is a regulatory switch in the interferon-mediated antiviral response

    Journal: Science Advances

    doi: 10.1126/sciadv.aef5520

    ( A ) Schematic type I IFN pathway representation. Created in BioRender. Paeschke, K. (2026) https://BioRender.com/5jo0emg . ( B ) Representative confocal images (left) and quantification (right) of G3BP1-positive SGs (green) and DAPI (blue). The Mann-Whitney U test compared N = 683 WT with N = 1060 DHX36 KO cells; each point is the image mean of multiple cells ± SD. ( C and D ) Western blot quantification of phosphor-PKR and phosphor-eIF2α levels in WT and DHX36 KO cells. Unpaired two-tailed t test on N = 5 independent experiments ± SD. Representative blots are shown in fig. S1 [(A) and (B)]. ( E ) Evaluation of G3BP1 foci/cells by IF staining of the indicated cell lines. The Mann-Whitney U test compared N = 379 (WT) with N = 505 (DHX36 KO), N = 420 (PKR KO), and N = 358 (DHX36/PKR KO); each point is the image mean of multiple cells ± SD. Analysis corresponds to representative images in fig. S1E. ( F ) Western blot quantification of the phosphor-eIF2α levels in the indicated cell lines. Unpaired two-tailed t test, N ≥ 5 independent experiments ± SD. ( G ) Evaluation of CFAs of WT and DHX36 KO cells. Representative CFA images can be found in fig. S1F. ( H and I ) Cell confluency measurements of WT and DHX36 KO cells. Data from N = 6 independent experiments ± SE. ( J ) Evaluation of G3BP1-positive cells upon poly(I:C) treatment for the indicated periods by IF. The Mann-Whitney U test compared N ≥ 838 WT cells with N ≥ 1050 DHX36 KO cells ± SD for each condition. Representative confocal microscopy images are shown in fig. S1J. ( K ) Same as in (C) but with poly(I:C)-treated cells; Data from N = 3 independent experiments. Representative Western blot is shown in fig. S1K. ( L ) Same as in (F) but with poly(I:C)-treated cells.
    Figure Legend Snippet: ( A ) Schematic type I IFN pathway representation. Created in BioRender. Paeschke, K. (2026) https://BioRender.com/5jo0emg . ( B ) Representative confocal images (left) and quantification (right) of G3BP1-positive SGs (green) and DAPI (blue). The Mann-Whitney U test compared N = 683 WT with N = 1060 DHX36 KO cells; each point is the image mean of multiple cells ± SD. ( C and D ) Western blot quantification of phosphor-PKR and phosphor-eIF2α levels in WT and DHX36 KO cells. Unpaired two-tailed t test on N = 5 independent experiments ± SD. Representative blots are shown in fig. S1 [(A) and (B)]. ( E ) Evaluation of G3BP1 foci/cells by IF staining of the indicated cell lines. The Mann-Whitney U test compared N = 379 (WT) with N = 505 (DHX36 KO), N = 420 (PKR KO), and N = 358 (DHX36/PKR KO); each point is the image mean of multiple cells ± SD. Analysis corresponds to representative images in fig. S1E. ( F ) Western blot quantification of the phosphor-eIF2α levels in the indicated cell lines. Unpaired two-tailed t test, N ≥ 5 independent experiments ± SD. ( G ) Evaluation of CFAs of WT and DHX36 KO cells. Representative CFA images can be found in fig. S1F. ( H and I ) Cell confluency measurements of WT and DHX36 KO cells. Data from N = 6 independent experiments ± SE. ( J ) Evaluation of G3BP1-positive cells upon poly(I:C) treatment for the indicated periods by IF. The Mann-Whitney U test compared N ≥ 838 WT cells with N ≥ 1050 DHX36 KO cells ± SD for each condition. Representative confocal microscopy images are shown in fig. S1J. ( K ) Same as in (C) but with poly(I:C)-treated cells; Data from N = 3 independent experiments. Representative Western blot is shown in fig. S1K. ( L ) Same as in (F) but with poly(I:C)-treated cells.

    Techniques Used: MANN-WHITNEY, Western Blot, Two Tailed Test, Staining, Confocal Microscopy

    ( A ) RNA-seq analysis of JAK/STAT pathway genes and ISGs of WT and DHX36 KO cells 24 hours post–poly(I:C) treatment. RNA-seq was performed on a HiSeq 2500 platform, comparing HEK293 WT and DHX36 KO cells. Genes were annotated to the hg19 human genome using TopHat2 and further processed using Cufflinks and Cuffdiff . Only genes with >5 Reads Per Kilobase Million (RPKM) were taken into account. Data from N = 3 independent experiments. ( B ) Type I IFN secretion of HEK293 WT and DHX36 KO cells, treated with poly(I:C) for 24 hours. The IFN concentrations were determined using THP1-Dual TBK1/IKKε/IKKα/IKKβ −/− reporter cell system and by comparison to an IFN-α standard curve. Data from N = 6 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( C to F ) Transcript levels of the ISGs IFNB1 , IFIT1 , RIG-I , and RSAD2 in HEK293 WT and DHX36 KO after poly(I:C) treatment for the indicated periods, assessed by qPCR. Data from at least N = 5 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( G ) Cumulative distribution function analysis reveals an increased abundance of DHX36 target mRNAs following 24 hours poly(I:C) treatment of HEK293 WT cells. DHX36 target mRNAs were binned on the basis of their binding intensities [Normalized Crosslinked Reads per Million (NXPM)] identified in DHX36 PAR-CLIP from Sauer et al. . Data from N = 3 independent experiments were evaluated using a two-sided Kolmogorov-Smirnov test. ( H ) Same as in (A), but comparing the abundance of DHX36 target mRNAs in DHX36 KO cells to WT cells. ( I ) Same as in (C), but comparing the abundance of DHX36 target mRNAs between DHX36 KO cells and 24-hour poly(I:C)-treated WT cells. logFC, log fold change.
    Figure Legend Snippet: ( A ) RNA-seq analysis of JAK/STAT pathway genes and ISGs of WT and DHX36 KO cells 24 hours post–poly(I:C) treatment. RNA-seq was performed on a HiSeq 2500 platform, comparing HEK293 WT and DHX36 KO cells. Genes were annotated to the hg19 human genome using TopHat2 and further processed using Cufflinks and Cuffdiff . Only genes with >5 Reads Per Kilobase Million (RPKM) were taken into account. Data from N = 3 independent experiments. ( B ) Type I IFN secretion of HEK293 WT and DHX36 KO cells, treated with poly(I:C) for 24 hours. The IFN concentrations were determined using THP1-Dual TBK1/IKKε/IKKα/IKKβ −/− reporter cell system and by comparison to an IFN-α standard curve. Data from N = 6 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( C to F ) Transcript levels of the ISGs IFNB1 , IFIT1 , RIG-I , and RSAD2 in HEK293 WT and DHX36 KO after poly(I:C) treatment for the indicated periods, assessed by qPCR. Data from at least N = 5 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( G ) Cumulative distribution function analysis reveals an increased abundance of DHX36 target mRNAs following 24 hours poly(I:C) treatment of HEK293 WT cells. DHX36 target mRNAs were binned on the basis of their binding intensities [Normalized Crosslinked Reads per Million (NXPM)] identified in DHX36 PAR-CLIP from Sauer et al. . Data from N = 3 independent experiments were evaluated using a two-sided Kolmogorov-Smirnov test. ( H ) Same as in (A), but comparing the abundance of DHX36 target mRNAs in DHX36 KO cells to WT cells. ( I ) Same as in (C), but comparing the abundance of DHX36 target mRNAs between DHX36 KO cells and 24-hour poly(I:C)-treated WT cells. logFC, log fold change.

    Techniques Used: RNA Sequencing, Comparison, Two Tailed Test, Binding Assay

    ( A ) Transcript levels of the ISGs IFIT1 , RIG-I , and RSAD2 , assessed by qPCR, after 24 hours of poly(I:C) treatment of HEK293 WT, DHX36 KO, and DHX36 OE cells. Data from N = 3 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( B ) Same as in (A), but comparing the ISG transcript levels in HEK293 WT, DHX36 KO, and catalytically dead DHX36_EA cells. Data from N = 3 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( C ) Luciferase assay on HEK293 WT, DHX36 KO, and DHX36 OE cells that were transfected by electroporation with an in vitro–transcribed YFV replicon containing an RLuc reporter allowing for replication quantification. DHX36 overexpression was induced with tetracycline (0.5 μg/ml) 24 hours before YFV replicon transfection. Data are normalized to the YFV levels 4 hours posttransfection. Data from N = 3 independent experiments were evaluated by the unpaired two-tailed t test and Bonferroni correction ± SD. ( D ) Statistical analysis of the melting temperature of recombinant DHX36 in the presence or absence of YFV replicon RNA or poly(I:C), determined by DSF. Corresponding thermal denaturation curves are shown in fig. S3F; exact T melt and Δ T melt values are listed in fig. S3G. Data represent N = 4 independent experiments, analyzed using the unpaired t test with Bonferroni correction; error bars indicate ± SD. ( E ) Representative gel of an EMSA showing the binding of recombinant DHX36 to YFV replicon RNA.
    Figure Legend Snippet: ( A ) Transcript levels of the ISGs IFIT1 , RIG-I , and RSAD2 , assessed by qPCR, after 24 hours of poly(I:C) treatment of HEK293 WT, DHX36 KO, and DHX36 OE cells. Data from N = 3 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( B ) Same as in (A), but comparing the ISG transcript levels in HEK293 WT, DHX36 KO, and catalytically dead DHX36_EA cells. Data from N = 3 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( C ) Luciferase assay on HEK293 WT, DHX36 KO, and DHX36 OE cells that were transfected by electroporation with an in vitro–transcribed YFV replicon containing an RLuc reporter allowing for replication quantification. DHX36 overexpression was induced with tetracycline (0.5 μg/ml) 24 hours before YFV replicon transfection. Data are normalized to the YFV levels 4 hours posttransfection. Data from N = 3 independent experiments were evaluated by the unpaired two-tailed t test and Bonferroni correction ± SD. ( D ) Statistical analysis of the melting temperature of recombinant DHX36 in the presence or absence of YFV replicon RNA or poly(I:C), determined by DSF. Corresponding thermal denaturation curves are shown in fig. S3F; exact T melt and Δ T melt values are listed in fig. S3G. Data represent N = 4 independent experiments, analyzed using the unpaired t test with Bonferroni correction; error bars indicate ± SD. ( E ) Representative gel of an EMSA showing the binding of recombinant DHX36 to YFV replicon RNA.

    Techniques Used: Two Tailed Test, Luciferase, Transfection, Electroporation, In Vitro, Over Expression, Recombinant, Binding Assay

    ( A ) Co-IP pull-downs of DHX36 performed under both untreated and poly(I:C)-treated conditions. N-terminally FLAG-tagged DHX36 was transiently expressed in HEK293 cells for Co-IP studies. After cell lysis, samples were treated with or without RNase A before pull-down using anti-FLAG beads. Western blots were evaluated using the anti-FLAG antibody for detection of DHX36 and antibodies against PKR and tubulin. The blots shown are representative of N = 3 independent experiments. Supporting data can be found in fig. S4A. ( B ) Western blot signal intensity quantification of phosphorylated p65 in HEK293 WT and DHX36 KO cells, nontreated or irradiated with UV light (20 J/m 2 ). Samples were taken 15, 30, or 60 min post–UV treatment. Signal intensity was normalized to p65 that was normalized to actin. Data from N = 3 independent experiments. Supporting data can be found in fig. S4B. ( C ) Fold induction of phosphorylated p65 in HEK293 WT and DHX36 KO cells, nontreated or poly(I:C) treated for 24 hours. Western blot signal intensity was normalized to actin and nontreated WT. Data from N = 5 independent experiments. ( D to F ) Transcript levels of TNF α, IL6 , and IL8 HEK293 WT and DHX36 KO cells, treated with poly(I:C) for 24 hours, assessed by qPCR. Data from at least N = 5 independent experiments ± SD. Statistical analysis evaluated using the Mann-Whitney U test with Bonferroni correction revealed no significant changes.
    Figure Legend Snippet: ( A ) Co-IP pull-downs of DHX36 performed under both untreated and poly(I:C)-treated conditions. N-terminally FLAG-tagged DHX36 was transiently expressed in HEK293 cells for Co-IP studies. After cell lysis, samples were treated with or without RNase A before pull-down using anti-FLAG beads. Western blots were evaluated using the anti-FLAG antibody for detection of DHX36 and antibodies against PKR and tubulin. The blots shown are representative of N = 3 independent experiments. Supporting data can be found in fig. S4A. ( B ) Western blot signal intensity quantification of phosphorylated p65 in HEK293 WT and DHX36 KO cells, nontreated or irradiated with UV light (20 J/m 2 ). Samples were taken 15, 30, or 60 min post–UV treatment. Signal intensity was normalized to p65 that was normalized to actin. Data from N = 3 independent experiments. Supporting data can be found in fig. S4B. ( C ) Fold induction of phosphorylated p65 in HEK293 WT and DHX36 KO cells, nontreated or poly(I:C) treated for 24 hours. Western blot signal intensity was normalized to actin and nontreated WT. Data from N = 5 independent experiments. ( D to F ) Transcript levels of TNF α, IL6 , and IL8 HEK293 WT and DHX36 KO cells, treated with poly(I:C) for 24 hours, assessed by qPCR. Data from at least N = 5 independent experiments ± SD. Statistical analysis evaluated using the Mann-Whitney U test with Bonferroni correction revealed no significant changes.

    Techniques Used: Co-Immunoprecipitation Assay, Lysis, Western Blot, Irradiation, MANN-WHITNEY

    ( A ) Transcript levels of IFNB1 in HEK293 WT, DHX36 KO, PKR KO, DHX36/PKR KO, RIG-I, and DHX36/RIG-I KO cells, assessed by qPCR after 24 hours of poly(I:C) treatment. Data from N = 6 independent experiments evaluated by the unpaired two-tailed t test with Bonferroni correction ± SD. ( B ) Schematic and simplified representation of the initial antiviral signal transduction focused on PKR and RIG-I. Created in BioRender. Paeschke, K. (2026) https://BioRender.com/1gkf0z7 . ( C to F ) Transcript levels of IFIT1 , RIG-I , ADAR1 , and IRF7 HEK293 WT, DHX36 KO, PKR KO, DHX36/PKR KO, RIG-I, and DHX36/RIG-I KO cells, assessed by qPCR 24 hours after poly(I:C) treatment. Data from at least N = 6 independent experiments evaluated by the unpaired two-tailed t test with Bonferroni correction ± SD.
    Figure Legend Snippet: ( A ) Transcript levels of IFNB1 in HEK293 WT, DHX36 KO, PKR KO, DHX36/PKR KO, RIG-I, and DHX36/RIG-I KO cells, assessed by qPCR after 24 hours of poly(I:C) treatment. Data from N = 6 independent experiments evaluated by the unpaired two-tailed t test with Bonferroni correction ± SD. ( B ) Schematic and simplified representation of the initial antiviral signal transduction focused on PKR and RIG-I. Created in BioRender. Paeschke, K. (2026) https://BioRender.com/1gkf0z7 . ( C to F ) Transcript levels of IFIT1 , RIG-I , ADAR1 , and IRF7 HEK293 WT, DHX36 KO, PKR KO, DHX36/PKR KO, RIG-I, and DHX36/RIG-I KO cells, assessed by qPCR 24 hours after poly(I:C) treatment. Data from at least N = 6 independent experiments evaluated by the unpaired two-tailed t test with Bonferroni correction ± SD.

    Techniques Used: Two Tailed Test, Transduction



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    ( A ) Schematic type I IFN pathway representation. Created in BioRender. Paeschke, K. (2026) https://BioRender.com/5jo0emg . ( B ) Representative confocal images (left) and quantification (right) of G3BP1-positive SGs (green) and DAPI (blue). The Mann-Whitney U test compared N = 683 WT with N = 1060 DHX36 KO cells; each point is the image mean of multiple cells ± SD. ( C and D ) Western blot quantification of phosphor-PKR and phosphor-eIF2α levels in WT and DHX36 KO cells. Unpaired two-tailed t test on N = 5 independent experiments ± SD. Representative blots are shown in fig. S1 [(A) and (B)]. ( E ) Evaluation of G3BP1 foci/cells by IF staining of the indicated cell lines. The Mann-Whitney U test compared N = 379 (WT) with N = 505 (DHX36 KO), N = 420 (PKR KO), and N = 358 (DHX36/PKR KO); each point is the image mean of multiple cells ± SD. Analysis corresponds to representative images in fig. S1E. ( F ) Western blot quantification of the phosphor-eIF2α levels in the indicated cell lines. Unpaired two-tailed t test, N ≥ 5 independent experiments ± SD. ( G ) Evaluation of CFAs of WT and DHX36 KO cells. Representative CFA images can be found in fig. S1F. ( H and I ) Cell confluency measurements of WT and DHX36 KO cells. Data from N = 6 independent experiments ± SE. ( J ) Evaluation of G3BP1-positive cells upon <t>poly(I:C)</t> treatment for the indicated periods by IF. The Mann-Whitney U test compared N ≥ 838 WT cells with N ≥ 1050 DHX36 KO cells ± SD for each condition. Representative confocal microscopy images are shown in fig. S1J. ( K ) Same as in (C) but with poly(I:C)-treated cells; Data from N = 3 independent experiments. Representative Western blot is shown in fig. S1K. ( L ) Same as in (F) but with poly(I:C)-treated cells.
    Cat Tlrl Pic Poly I C Low Molecular Weight Invivogen Cat Tlrl Picw, supplied by InvivoGen, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ( A ) Schematic type I IFN pathway representation. Created in BioRender. Paeschke, K. (2026) https://BioRender.com/5jo0emg . ( B ) Representative confocal images (left) and quantification (right) of G3BP1-positive SGs (green) and DAPI (blue). The Mann-Whitney U test compared N = 683 WT with N = 1060 DHX36 KO cells; each point is the image mean of multiple cells ± SD. ( C and D ) Western blot quantification of phosphor-PKR and phosphor-eIF2α levels in WT and DHX36 KO cells. Unpaired two-tailed t test on N = 5 independent experiments ± SD. Representative blots are shown in fig. S1 [(A) and (B)]. ( E ) Evaluation of G3BP1 foci/cells by IF staining of the indicated cell lines. The Mann-Whitney U test compared N = 379 (WT) with N = 505 (DHX36 KO), N = 420 (PKR KO), and N = 358 (DHX36/PKR KO); each point is the image mean of multiple cells ± SD. Analysis corresponds to representative images in fig. S1E. ( F ) Western blot quantification of the phosphor-eIF2α levels in the indicated cell lines. Unpaired two-tailed t test, N ≥ 5 independent experiments ± SD. ( G ) Evaluation of CFAs of WT and DHX36 KO cells. Representative CFA images can be found in fig. S1F. ( H and I ) Cell confluency measurements of WT and DHX36 KO cells. Data from N = 6 independent experiments ± SE. ( J ) Evaluation of G3BP1-positive cells upon <t>poly(I:C)</t> treatment for the indicated periods by IF. The Mann-Whitney U test compared N ≥ 838 WT cells with N ≥ 1050 DHX36 KO cells ± SD for each condition. Representative confocal microscopy images are shown in fig. S1J. ( K ) Same as in (C) but with poly(I:C)-treated cells; Data from N = 3 independent experiments. Representative Western blot is shown in fig. S1K. ( L ) Same as in (F) but with poly(I:C)-treated cells.
    Dsrna Analog Poly I C Trlr Pic, supplied by InvivoGen, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    ( A ) Schematic type I IFN pathway representation. Created in BioRender. Paeschke, K. (2026) https://BioRender.com/5jo0emg . ( B ) Representative confocal images (left) and quantification (right) of G3BP1-positive SGs (green) and DAPI (blue). The Mann-Whitney U test compared N = 683 WT with N = 1060 DHX36 KO cells; each point is the image mean of multiple cells ± SD. ( C and D ) Western blot quantification of phosphor-PKR and phosphor-eIF2α levels in WT and DHX36 KO cells. Unpaired two-tailed t test on N = 5 independent experiments ± SD. Representative blots are shown in fig. S1 [(A) and (B)]. ( E ) Evaluation of G3BP1 foci/cells by IF staining of the indicated cell lines. The Mann-Whitney U test compared N = 379 (WT) with N = 505 (DHX36 KO), N = 420 (PKR KO), and N = 358 (DHX36/PKR KO); each point is the image mean of multiple cells ± SD. Analysis corresponds to representative images in fig. S1E. ( F ) Western blot quantification of the phosphor-eIF2α levels in the indicated cell lines. Unpaired two-tailed t test, N ≥ 5 independent experiments ± SD. ( G ) Evaluation of CFAs of WT and DHX36 KO cells. Representative CFA images can be found in fig. S1F. ( H and I ) Cell confluency measurements of WT and DHX36 KO cells. Data from N = 6 independent experiments ± SE. ( J ) Evaluation of G3BP1-positive cells upon poly(I:C) treatment for the indicated periods by IF. The Mann-Whitney U test compared N ≥ 838 WT cells with N ≥ 1050 DHX36 KO cells ± SD for each condition. Representative confocal microscopy images are shown in fig. S1J. ( K ) Same as in (C) but with poly(I:C)-treated cells; Data from N = 3 independent experiments. Representative Western blot is shown in fig. S1K. ( L ) Same as in (F) but with poly(I:C)-treated cells.

    Journal: Science Advances

    Article Title: DHX36 is a regulatory switch in the interferon-mediated antiviral response

    doi: 10.1126/sciadv.aef5520

    Figure Lengend Snippet: ( A ) Schematic type I IFN pathway representation. Created in BioRender. Paeschke, K. (2026) https://BioRender.com/5jo0emg . ( B ) Representative confocal images (left) and quantification (right) of G3BP1-positive SGs (green) and DAPI (blue). The Mann-Whitney U test compared N = 683 WT with N = 1060 DHX36 KO cells; each point is the image mean of multiple cells ± SD. ( C and D ) Western blot quantification of phosphor-PKR and phosphor-eIF2α levels in WT and DHX36 KO cells. Unpaired two-tailed t test on N = 5 independent experiments ± SD. Representative blots are shown in fig. S1 [(A) and (B)]. ( E ) Evaluation of G3BP1 foci/cells by IF staining of the indicated cell lines. The Mann-Whitney U test compared N = 379 (WT) with N = 505 (DHX36 KO), N = 420 (PKR KO), and N = 358 (DHX36/PKR KO); each point is the image mean of multiple cells ± SD. Analysis corresponds to representative images in fig. S1E. ( F ) Western blot quantification of the phosphor-eIF2α levels in the indicated cell lines. Unpaired two-tailed t test, N ≥ 5 independent experiments ± SD. ( G ) Evaluation of CFAs of WT and DHX36 KO cells. Representative CFA images can be found in fig. S1F. ( H and I ) Cell confluency measurements of WT and DHX36 KO cells. Data from N = 6 independent experiments ± SE. ( J ) Evaluation of G3BP1-positive cells upon poly(I:C) treatment for the indicated periods by IF. The Mann-Whitney U test compared N ≥ 838 WT cells with N ≥ 1050 DHX36 KO cells ± SD for each condition. Representative confocal microscopy images are shown in fig. S1J. ( K ) Same as in (C) but with poly(I:C)-treated cells; Data from N = 3 independent experiments. Representative Western blot is shown in fig. S1K. ( L ) Same as in (F) but with poly(I:C)-treated cells.

    Article Snippet: Poly(I:C) treatments were performed using poly(I:C) (1 μg ml −1 ; InvivoGen; tlrl-pic) delivered by transfection with Lipofectamine 2000 (Thermo Fisher Scientific) according to the manufacturer’s protocol.

    Techniques: MANN-WHITNEY, Western Blot, Two Tailed Test, Staining, Confocal Microscopy

    ( A ) RNA-seq analysis of JAK/STAT pathway genes and ISGs of WT and DHX36 KO cells 24 hours post–poly(I:C) treatment. RNA-seq was performed on a HiSeq 2500 platform, comparing HEK293 WT and DHX36 KO cells. Genes were annotated to the hg19 human genome using TopHat2 and further processed using Cufflinks and Cuffdiff . Only genes with >5 Reads Per Kilobase Million (RPKM) were taken into account. Data from N = 3 independent experiments. ( B ) Type I IFN secretion of HEK293 WT and DHX36 KO cells, treated with poly(I:C) for 24 hours. The IFN concentrations were determined using THP1-Dual TBK1/IKKε/IKKα/IKKβ −/− reporter cell system and by comparison to an IFN-α standard curve. Data from N = 6 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( C to F ) Transcript levels of the ISGs IFNB1 , IFIT1 , RIG-I , and RSAD2 in HEK293 WT and DHX36 KO after poly(I:C) treatment for the indicated periods, assessed by qPCR. Data from at least N = 5 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( G ) Cumulative distribution function analysis reveals an increased abundance of DHX36 target mRNAs following 24 hours poly(I:C) treatment of HEK293 WT cells. DHX36 target mRNAs were binned on the basis of their binding intensities [Normalized Crosslinked Reads per Million (NXPM)] identified in DHX36 PAR-CLIP from Sauer et al. . Data from N = 3 independent experiments were evaluated using a two-sided Kolmogorov-Smirnov test. ( H ) Same as in (A), but comparing the abundance of DHX36 target mRNAs in DHX36 KO cells to WT cells. ( I ) Same as in (C), but comparing the abundance of DHX36 target mRNAs between DHX36 KO cells and 24-hour poly(I:C)-treated WT cells. logFC, log fold change.

    Journal: Science Advances

    Article Title: DHX36 is a regulatory switch in the interferon-mediated antiviral response

    doi: 10.1126/sciadv.aef5520

    Figure Lengend Snippet: ( A ) RNA-seq analysis of JAK/STAT pathway genes and ISGs of WT and DHX36 KO cells 24 hours post–poly(I:C) treatment. RNA-seq was performed on a HiSeq 2500 platform, comparing HEK293 WT and DHX36 KO cells. Genes were annotated to the hg19 human genome using TopHat2 and further processed using Cufflinks and Cuffdiff . Only genes with >5 Reads Per Kilobase Million (RPKM) were taken into account. Data from N = 3 independent experiments. ( B ) Type I IFN secretion of HEK293 WT and DHX36 KO cells, treated with poly(I:C) for 24 hours. The IFN concentrations were determined using THP1-Dual TBK1/IKKε/IKKα/IKKβ −/− reporter cell system and by comparison to an IFN-α standard curve. Data from N = 6 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( C to F ) Transcript levels of the ISGs IFNB1 , IFIT1 , RIG-I , and RSAD2 in HEK293 WT and DHX36 KO after poly(I:C) treatment for the indicated periods, assessed by qPCR. Data from at least N = 5 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( G ) Cumulative distribution function analysis reveals an increased abundance of DHX36 target mRNAs following 24 hours poly(I:C) treatment of HEK293 WT cells. DHX36 target mRNAs were binned on the basis of their binding intensities [Normalized Crosslinked Reads per Million (NXPM)] identified in DHX36 PAR-CLIP from Sauer et al. . Data from N = 3 independent experiments were evaluated using a two-sided Kolmogorov-Smirnov test. ( H ) Same as in (A), but comparing the abundance of DHX36 target mRNAs in DHX36 KO cells to WT cells. ( I ) Same as in (C), but comparing the abundance of DHX36 target mRNAs between DHX36 KO cells and 24-hour poly(I:C)-treated WT cells. logFC, log fold change.

    Article Snippet: Poly(I:C) treatments were performed using poly(I:C) (1 μg ml −1 ; InvivoGen; tlrl-pic) delivered by transfection with Lipofectamine 2000 (Thermo Fisher Scientific) according to the manufacturer’s protocol.

    Techniques: RNA Sequencing, Comparison, Two Tailed Test, Binding Assay

    ( A ) Transcript levels of the ISGs IFIT1 , RIG-I , and RSAD2 , assessed by qPCR, after 24 hours of poly(I:C) treatment of HEK293 WT, DHX36 KO, and DHX36 OE cells. Data from N = 3 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( B ) Same as in (A), but comparing the ISG transcript levels in HEK293 WT, DHX36 KO, and catalytically dead DHX36_EA cells. Data from N = 3 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( C ) Luciferase assay on HEK293 WT, DHX36 KO, and DHX36 OE cells that were transfected by electroporation with an in vitro–transcribed YFV replicon containing an RLuc reporter allowing for replication quantification. DHX36 overexpression was induced with tetracycline (0.5 μg/ml) 24 hours before YFV replicon transfection. Data are normalized to the YFV levels 4 hours posttransfection. Data from N = 3 independent experiments were evaluated by the unpaired two-tailed t test and Bonferroni correction ± SD. ( D ) Statistical analysis of the melting temperature of recombinant DHX36 in the presence or absence of YFV replicon RNA or poly(I:C), determined by DSF. Corresponding thermal denaturation curves are shown in fig. S3F; exact T melt and Δ T melt values are listed in fig. S3G. Data represent N = 4 independent experiments, analyzed using the unpaired t test with Bonferroni correction; error bars indicate ± SD. ( E ) Representative gel of an EMSA showing the binding of recombinant DHX36 to YFV replicon RNA.

    Journal: Science Advances

    Article Title: DHX36 is a regulatory switch in the interferon-mediated antiviral response

    doi: 10.1126/sciadv.aef5520

    Figure Lengend Snippet: ( A ) Transcript levels of the ISGs IFIT1 , RIG-I , and RSAD2 , assessed by qPCR, after 24 hours of poly(I:C) treatment of HEK293 WT, DHX36 KO, and DHX36 OE cells. Data from N = 3 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( B ) Same as in (A), but comparing the ISG transcript levels in HEK293 WT, DHX36 KO, and catalytically dead DHX36_EA cells. Data from N = 3 independent experiments evaluated by the unpaired two-tailed t test ± SD. ( C ) Luciferase assay on HEK293 WT, DHX36 KO, and DHX36 OE cells that were transfected by electroporation with an in vitro–transcribed YFV replicon containing an RLuc reporter allowing for replication quantification. DHX36 overexpression was induced with tetracycline (0.5 μg/ml) 24 hours before YFV replicon transfection. Data are normalized to the YFV levels 4 hours posttransfection. Data from N = 3 independent experiments were evaluated by the unpaired two-tailed t test and Bonferroni correction ± SD. ( D ) Statistical analysis of the melting temperature of recombinant DHX36 in the presence or absence of YFV replicon RNA or poly(I:C), determined by DSF. Corresponding thermal denaturation curves are shown in fig. S3F; exact T melt and Δ T melt values are listed in fig. S3G. Data represent N = 4 independent experiments, analyzed using the unpaired t test with Bonferroni correction; error bars indicate ± SD. ( E ) Representative gel of an EMSA showing the binding of recombinant DHX36 to YFV replicon RNA.

    Article Snippet: Poly(I:C) treatments were performed using poly(I:C) (1 μg ml −1 ; InvivoGen; tlrl-pic) delivered by transfection with Lipofectamine 2000 (Thermo Fisher Scientific) according to the manufacturer’s protocol.

    Techniques: Two Tailed Test, Luciferase, Transfection, Electroporation, In Vitro, Over Expression, Recombinant, Binding Assay

    ( A ) Co-IP pull-downs of DHX36 performed under both untreated and poly(I:C)-treated conditions. N-terminally FLAG-tagged DHX36 was transiently expressed in HEK293 cells for Co-IP studies. After cell lysis, samples were treated with or without RNase A before pull-down using anti-FLAG beads. Western blots were evaluated using the anti-FLAG antibody for detection of DHX36 and antibodies against PKR and tubulin. The blots shown are representative of N = 3 independent experiments. Supporting data can be found in fig. S4A. ( B ) Western blot signal intensity quantification of phosphorylated p65 in HEK293 WT and DHX36 KO cells, nontreated or irradiated with UV light (20 J/m 2 ). Samples were taken 15, 30, or 60 min post–UV treatment. Signal intensity was normalized to p65 that was normalized to actin. Data from N = 3 independent experiments. Supporting data can be found in fig. S4B. ( C ) Fold induction of phosphorylated p65 in HEK293 WT and DHX36 KO cells, nontreated or poly(I:C) treated for 24 hours. Western blot signal intensity was normalized to actin and nontreated WT. Data from N = 5 independent experiments. ( D to F ) Transcript levels of TNF α, IL6 , and IL8 HEK293 WT and DHX36 KO cells, treated with poly(I:C) for 24 hours, assessed by qPCR. Data from at least N = 5 independent experiments ± SD. Statistical analysis evaluated using the Mann-Whitney U test with Bonferroni correction revealed no significant changes.

    Journal: Science Advances

    Article Title: DHX36 is a regulatory switch in the interferon-mediated antiviral response

    doi: 10.1126/sciadv.aef5520

    Figure Lengend Snippet: ( A ) Co-IP pull-downs of DHX36 performed under both untreated and poly(I:C)-treated conditions. N-terminally FLAG-tagged DHX36 was transiently expressed in HEK293 cells for Co-IP studies. After cell lysis, samples were treated with or without RNase A before pull-down using anti-FLAG beads. Western blots were evaluated using the anti-FLAG antibody for detection of DHX36 and antibodies against PKR and tubulin. The blots shown are representative of N = 3 independent experiments. Supporting data can be found in fig. S4A. ( B ) Western blot signal intensity quantification of phosphorylated p65 in HEK293 WT and DHX36 KO cells, nontreated or irradiated with UV light (20 J/m 2 ). Samples were taken 15, 30, or 60 min post–UV treatment. Signal intensity was normalized to p65 that was normalized to actin. Data from N = 3 independent experiments. Supporting data can be found in fig. S4B. ( C ) Fold induction of phosphorylated p65 in HEK293 WT and DHX36 KO cells, nontreated or poly(I:C) treated for 24 hours. Western blot signal intensity was normalized to actin and nontreated WT. Data from N = 5 independent experiments. ( D to F ) Transcript levels of TNF α, IL6 , and IL8 HEK293 WT and DHX36 KO cells, treated with poly(I:C) for 24 hours, assessed by qPCR. Data from at least N = 5 independent experiments ± SD. Statistical analysis evaluated using the Mann-Whitney U test with Bonferroni correction revealed no significant changes.

    Article Snippet: Poly(I:C) treatments were performed using poly(I:C) (1 μg ml −1 ; InvivoGen; tlrl-pic) delivered by transfection with Lipofectamine 2000 (Thermo Fisher Scientific) according to the manufacturer’s protocol.

    Techniques: Co-Immunoprecipitation Assay, Lysis, Western Blot, Irradiation, MANN-WHITNEY

    ( A ) Transcript levels of IFNB1 in HEK293 WT, DHX36 KO, PKR KO, DHX36/PKR KO, RIG-I, and DHX36/RIG-I KO cells, assessed by qPCR after 24 hours of poly(I:C) treatment. Data from N = 6 independent experiments evaluated by the unpaired two-tailed t test with Bonferroni correction ± SD. ( B ) Schematic and simplified representation of the initial antiviral signal transduction focused on PKR and RIG-I. Created in BioRender. Paeschke, K. (2026) https://BioRender.com/1gkf0z7 . ( C to F ) Transcript levels of IFIT1 , RIG-I , ADAR1 , and IRF7 HEK293 WT, DHX36 KO, PKR KO, DHX36/PKR KO, RIG-I, and DHX36/RIG-I KO cells, assessed by qPCR 24 hours after poly(I:C) treatment. Data from at least N = 6 independent experiments evaluated by the unpaired two-tailed t test with Bonferroni correction ± SD.

    Journal: Science Advances

    Article Title: DHX36 is a regulatory switch in the interferon-mediated antiviral response

    doi: 10.1126/sciadv.aef5520

    Figure Lengend Snippet: ( A ) Transcript levels of IFNB1 in HEK293 WT, DHX36 KO, PKR KO, DHX36/PKR KO, RIG-I, and DHX36/RIG-I KO cells, assessed by qPCR after 24 hours of poly(I:C) treatment. Data from N = 6 independent experiments evaluated by the unpaired two-tailed t test with Bonferroni correction ± SD. ( B ) Schematic and simplified representation of the initial antiviral signal transduction focused on PKR and RIG-I. Created in BioRender. Paeschke, K. (2026) https://BioRender.com/1gkf0z7 . ( C to F ) Transcript levels of IFIT1 , RIG-I , ADAR1 , and IRF7 HEK293 WT, DHX36 KO, PKR KO, DHX36/PKR KO, RIG-I, and DHX36/RIG-I KO cells, assessed by qPCR 24 hours after poly(I:C) treatment. Data from at least N = 6 independent experiments evaluated by the unpaired two-tailed t test with Bonferroni correction ± SD.

    Article Snippet: Poly(I:C) treatments were performed using poly(I:C) (1 μg ml −1 ; InvivoGen; tlrl-pic) delivered by transfection with Lipofectamine 2000 (Thermo Fisher Scientific) according to the manufacturer’s protocol.

    Techniques: Two Tailed Test, Transduction