ucyn a whole genome diel transcription patterns  (ATCC)


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    ATCC ucyn a whole genome diel transcription patterns
    Transcription of genes for replication and cell division in <t>UCYN-A.</t> (A) (Upper panel) Diel transcription patterns for cell division and replication genes in UCYN-A1 and UCYN-A2 over the light-dark cycle. Hierarchical clustering of genes was based on Pearson correlations between their transcription profiles. The transcription values of the genes were standardized at each time point, and the blue-red scale shows by how many standard deviations a transcription value was lower (blue) or higher (red) than the mean transcription values over the diel cycle (Z score). The Gene ID and the gene product corresponding to each gene for UCYN-A1 and UCYN-A2 are shown. Time periods are indicated on the x axis as “L” (light) and “D” (dark) followed by a number corresponding to the hour after the sunrise and sunset periods started. The second light-dark cycle is indicated as “2D” followed by a number corresponding to the hour of entry into the light or dark period. (Lower panel) Epifluorescence micrographs of dividing UCYN-A2 detected with CARD-FISH ( 19 ). (B) Two big clusters of UCYN-A2 cells and the attached haptophyte host. (Left panel) The nucleus of the host and the UCYN-A2 cells were visualized with DAPI stain (blue). (Right panel) The UCYN-A2 strain (red) and its haptophyte host (green). (C) Two different associations of UCYN-A2 with its haptophyte dividing in samples from Scripps Pier.
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    Images

    1) Product Images from "The Transcriptional Cycle Is Suited to Daytime N2 Fixation in the Unicellular Cyanobacterium “Candidatus Atelocyanobacterium thalassa” (UCYN-A)"

    Article Title: The Transcriptional Cycle Is Suited to Daytime N2 Fixation in the Unicellular Cyanobacterium “Candidatus Atelocyanobacterium thalassa” (UCYN-A)

    Journal: mBio

    doi: 10.1128/mBio.02495-18

    Transcription of genes for replication and cell division in UCYN-A. (A) (Upper panel) Diel transcription patterns for cell division and replication genes in UCYN-A1 and UCYN-A2 over the light-dark cycle. Hierarchical clustering of genes was based on Pearson correlations between their transcription profiles. The transcription values of the genes were standardized at each time point, and the blue-red scale shows by how many standard deviations a transcription value was lower (blue) or higher (red) than the mean transcription values over the diel cycle (Z score). The Gene ID and the gene product corresponding to each gene for UCYN-A1 and UCYN-A2 are shown. Time periods are indicated on the x axis as “L” (light) and “D” (dark) followed by a number corresponding to the hour after the sunrise and sunset periods started. The second light-dark cycle is indicated as “2D” followed by a number corresponding to the hour of entry into the light or dark period. (Lower panel) Epifluorescence micrographs of dividing UCYN-A2 detected with CARD-FISH ( 19 ). (B) Two big clusters of UCYN-A2 cells and the attached haptophyte host. (Left panel) The nucleus of the host and the UCYN-A2 cells were visualized with DAPI stain (blue). (Right panel) The UCYN-A2 strain (red) and its haptophyte host (green). (C) Two different associations of UCYN-A2 with its haptophyte dividing in samples from Scripps Pier.
    Figure Legend Snippet: Transcription of genes for replication and cell division in UCYN-A. (A) (Upper panel) Diel transcription patterns for cell division and replication genes in UCYN-A1 and UCYN-A2 over the light-dark cycle. Hierarchical clustering of genes was based on Pearson correlations between their transcription profiles. The transcription values of the genes were standardized at each time point, and the blue-red scale shows by how many standard deviations a transcription value was lower (blue) or higher (red) than the mean transcription values over the diel cycle (Z score). The Gene ID and the gene product corresponding to each gene for UCYN-A1 and UCYN-A2 are shown. Time periods are indicated on the x axis as “L” (light) and “D” (dark) followed by a number corresponding to the hour after the sunrise and sunset periods started. The second light-dark cycle is indicated as “2D” followed by a number corresponding to the hour of entry into the light or dark period. (Lower panel) Epifluorescence micrographs of dividing UCYN-A2 detected with CARD-FISH ( 19 ). (B) Two big clusters of UCYN-A2 cells and the attached haptophyte host. (Left panel) The nucleus of the host and the UCYN-A2 cells were visualized with DAPI stain (blue). (Right panel) The UCYN-A2 strain (red) and its haptophyte host (green). (C) Two different associations of UCYN-A2 with its haptophyte dividing in samples from Scripps Pier.

    Techniques Used: Fluorescence In Situ Hybridization, Staining

    Schematic model of UCYN-A showing the possible main cellular functions, metabolic pathways, and transporters.
    Figure Legend Snippet: Schematic model of UCYN-A showing the possible main cellular functions, metabolic pathways, and transporters.

    Techniques Used:

    Network showing the Pearson correlation for gene transcriptions in the unicellular N 2 -fixing cyanobacteria Cyanothece sp. ATCC 51142 ( Cyanothece ), C. watsonii WH 8501 ( Crocosphaera ), and UCYN-A. Shown here are key genes in major metabolic pathways with distinct diel transcription patterns. The genes are shown as connected if their correlation coefficient for transcription patterns is higher than 0.2. PPP, pentose phosphate pathway; PSI, photosystem I.
    Figure Legend Snippet: Network showing the Pearson correlation for gene transcriptions in the unicellular N 2 -fixing cyanobacteria Cyanothece sp. ATCC 51142 ( Cyanothece ), C. watsonii WH 8501 ( Crocosphaera ), and UCYN-A. Shown here are key genes in major metabolic pathways with distinct diel transcription patterns. The genes are shown as connected if their correlation coefficient for transcription patterns is higher than 0.2. PPP, pentose phosphate pathway; PSI, photosystem I.

    Techniques Used:

    2) Product Images from "Characterization of the Antibody Response to the Receptor Binding Domain of Botulinum Neurotoxin Serotypes A and E"

    Article Title: Characterization of the Antibody Response to the Receptor Binding Domain of Botulinum Neurotoxin Serotypes A and E

    Journal: Infection and Immunity

    doi: 10.1128/IAI.73.10.6998-7005.2005

    Protein modeling of HCR/A2 (Kyoto F) and HCR/E (Beluga). (A) Using the structures of BoNT/A (pdb:3bta), BoNT/B (pdb:1epw), and tetanus HCR (pdb:1doh) as templates, the three-dimensional structures of HCR/A2 and HCR/E B were generated using Swiss-Model. Ribbon diagrams of HCR/A1 (blue), HCR/A2 (red), and HCR/E B (black) are displayed in the upper panels. Molecular surface electrostatic potentials of each protein were computed using the Coulomb method and are displayed in the lower panels (blue, positive charge; red, negative charge; white, neutral). The regions of lowest structural homology between HCR/A1, HCRA2, and HCR/E are circled and labeled 1 through 4. (B) Enlarged view of region 5 highlighting the primary residues contributing to the electrostatic surface of the molecule (left panel). Sequence alignment of the peptides forming this region are displayed on the right with conserved charge residues highlighted.
    Figure Legend Snippet: Protein modeling of HCR/A2 (Kyoto F) and HCR/E (Beluga). (A) Using the structures of BoNT/A (pdb:3bta), BoNT/B (pdb:1epw), and tetanus HCR (pdb:1doh) as templates, the three-dimensional structures of HCR/A2 and HCR/E B were generated using Swiss-Model. Ribbon diagrams of HCR/A1 (blue), HCR/A2 (red), and HCR/E B (black) are displayed in the upper panels. Molecular surface electrostatic potentials of each protein were computed using the Coulomb method and are displayed in the lower panels (blue, positive charge; red, negative charge; white, neutral). The regions of lowest structural homology between HCR/A1, HCRA2, and HCR/E are circled and labeled 1 through 4. (B) Enlarged view of region 5 highlighting the primary residues contributing to the electrostatic surface of the molecule (left panel). Sequence alignment of the peptides forming this region are displayed on the right with conserved charge residues highlighted.

    Techniques Used: Generated, Labeling, Sequencing

    3) Product Images from "Combination Therapy Using Monoclonal Antibodies against Respiratory Syncytial Virus (RSV) G Glycoprotein Protects from RSV Disease in BALB/c Mice"

    Article Title: Combination Therapy Using Monoclonal Antibodies against Respiratory Syncytial Virus (RSV) G Glycoprotein Protects from RSV Disease in BALB/c Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0051485

    Antibody inhibition of leukocyte migration by mAbs 130-6D and 131-2G. The inhibition of RSV A2 G protein – mediated leukocyte chemotaxis by anti-RSV G antibodies (130-6D and 131-2G) was determined as described in Materials and Methods. Antibody was included in the lower chamber of the chemotaxis chamber, along with purified RSV A2 G protein, at an equal concentration (1X) (22.5 µg/mL) to that of purified RSV G protein (22.5 µg/mL) up to a 50-fold concentration (50X) of mAb to purified RSV G protein. For combination mAb inhibition, 1X and 50X of each antibody were used. Each condition was assayed three times per experiment. Data is presented as an average percent inhibition of RSV A2 G protein mediated leukocyte chemotaxis ± standard deviation relative to the amount of chemotaxis induced by RSV A2 G protein alone. Asterisk indicates significant difference in inhibition (p
    Figure Legend Snippet: Antibody inhibition of leukocyte migration by mAbs 130-6D and 131-2G. The inhibition of RSV A2 G protein – mediated leukocyte chemotaxis by anti-RSV G antibodies (130-6D and 131-2G) was determined as described in Materials and Methods. Antibody was included in the lower chamber of the chemotaxis chamber, along with purified RSV A2 G protein, at an equal concentration (1X) (22.5 µg/mL) to that of purified RSV G protein (22.5 µg/mL) up to a 50-fold concentration (50X) of mAb to purified RSV G protein. For combination mAb inhibition, 1X and 50X of each antibody were used. Each condition was assayed three times per experiment. Data is presented as an average percent inhibition of RSV A2 G protein mediated leukocyte chemotaxis ± standard deviation relative to the amount of chemotaxis induced by RSV A2 G protein alone. Asterisk indicates significant difference in inhibition (p

    Techniques Used: Inhibition, Migration, Chemotaxis Assay, Purification, Concentration Assay, Standard Deviation

    4) Product Images from "Diagnostic utility of LunX mRNA in peripheral blood and pleural fluid in patients with primary non-small cell lung cancer"

    Article Title: Diagnostic utility of LunX mRNA in peripheral blood and pleural fluid in patients with primary non-small cell lung cancer

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-8-156

    LunX mRNA is the most specific gene marker for lung cancer cells in peripheral blood . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin in all samples. For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as 50%, the median cannot be shown. " title="LunX mRNA is the most specific gene marker for ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: LunX mRNA is the most specific gene marker for lung cancer cells in peripheral blood . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin in all samples. For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as " --- " in each group. Where the frequency of negative cases is > 50%, the median cannot be shown.

    Techniques Used: Marker, Quantitative RT-PCR

    Expression of LunX mRNA in peripheral blood decreases shortly following the treatment of NSCLC . Peripheral blood samples from 12 NSCLC patients were collected 1 day before and 7 days after treatment as shown in Table 2. LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods.
    Figure Legend Snippet: Expression of LunX mRNA in peripheral blood decreases shortly following the treatment of NSCLC . Peripheral blood samples from 12 NSCLC patients were collected 1 day before and 7 days after treatment as shown in Table 2. LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. "Tb" represents 1 day before treatment and "Ta" represents 7 days after treatment. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The Wilcoxon Signed Ranks Test was used to analyze the gene expression levels before and after clinical treatment. P

    Techniques Used: Expressing, Quantitative RT-PCR, Marker

    Expression level of LunX mRNA in peripheral blood is correlated with the pathologic stage of NSCLC . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as 50%, the median cannot be shown. The K Independent Samples Test (Media Test) was used to analyze gene expression levels among NSCLC patients at different pathologic stages. P " title="Expression level of LunX mRNA in peripheral blood is correlated with the ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Expression level of LunX mRNA in peripheral blood is correlated with the pathologic stage of NSCLC . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as " --- " in each group. Where the frequency of negative cases is > 50%, the median cannot be shown. The K Independent Samples Test (Media Test) was used to analyze gene expression levels among NSCLC patients at different pathologic stages. P

    Techniques Used: Expressing, Quantitative RT-PCR, Marker

    LunX mRNA is the most specific gene marker with high sensitivity for NSCLC cells in pleural fluid . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each pleural fluid sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as 50%, the median cannot be shown. The Mann-Whitney U Test was used to compare the gene expression levels between NSCLC and tuberculo pleurisy groups. P " title="LunX mRNA is the most specific gene marker with ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: LunX mRNA is the most specific gene marker with high sensitivity for NSCLC cells in pleural fluid . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each pleural fluid sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as " --- " in each group. Where the frequency of negative cases is > 50%, the median cannot be shown. The Mann-Whitney U Test was used to compare the gene expression levels between NSCLC and tuberculo pleurisy groups. P

    Techniques Used: Marker, Quantitative RT-PCR, MANN-WHITNEY, Expressing

    5) Product Images from "Diagnostic utility of LunX mRNA in peripheral blood and pleural fluid in patients with primary non-small cell lung cancer"

    Article Title: Diagnostic utility of LunX mRNA in peripheral blood and pleural fluid in patients with primary non-small cell lung cancer

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-8-156

    LunX mRNA is the most specific gene marker for lung cancer cells in peripheral blood . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin in all samples. For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as 50%, the median cannot be shown. " title="... in peripheral blood . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: LunX mRNA is the most specific gene marker for lung cancer cells in peripheral blood . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin in all samples. For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as " --- " in each group. Where the frequency of negative cases is > 50%, the median cannot be shown.

    Techniques Used: Marker, Quantitative RT-PCR

    Expression of LunX mRNA in peripheral blood decreases shortly following the treatment of NSCLC . Peripheral blood samples from 12 NSCLC patients were collected 1 day before and 7 days after treatment as shown in Table 2. LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods.
    Figure Legend Snippet: Expression of LunX mRNA in peripheral blood decreases shortly following the treatment of NSCLC . Peripheral blood samples from 12 NSCLC patients were collected 1 day before and 7 days after treatment as shown in Table 2. LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. "Tb" represents 1 day before treatment and "Ta" represents 7 days after treatment. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The Wilcoxon Signed Ranks Test was used to analyze the gene expression levels before and after clinical treatment. P

    Techniques Used: Expressing, Quantitative RT-PCR, Marker

    Expression level of LunX mRNA in peripheral blood is correlated with the pathologic stage of NSCLC . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as 50%, the median cannot be shown. The K Independent Samples Test (Media Test) was used to analyze gene expression levels among NSCLC patients at different pathologic stages. P " title="... stage of NSCLC . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Expression level of LunX mRNA in peripheral blood is correlated with the pathologic stage of NSCLC . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as " --- " in each group. Where the frequency of negative cases is > 50%, the median cannot be shown. The K Independent Samples Test (Media Test) was used to analyze gene expression levels among NSCLC patients at different pathologic stages. P

    Techniques Used: Expressing, Quantitative RT-PCR, Marker

    LunX mRNA is the most specific gene marker with high sensitivity for NSCLC cells in pleural fluid . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each pleural fluid sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as 50%, the median cannot be shown. The Mann-Whitney U Test was used to compare the gene expression levels between NSCLC and tuberculo pleurisy groups. P " title="... in pleural fluid . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each pleural fluid ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: LunX mRNA is the most specific gene marker with high sensitivity for NSCLC cells in pleural fluid . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each pleural fluid sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as " --- " in each group. Where the frequency of negative cases is > 50%, the median cannot be shown. The Mann-Whitney U Test was used to compare the gene expression levels between NSCLC and tuberculo pleurisy groups. P

    Techniques Used: Marker, Quantitative RT-PCR, MANN-WHITNEY, Expressing

    6) Product Images from "Analysis of the Lactobacillus casei supragenome and its influence in species evolution and lifestyle adaptation"

    Article Title: Analysis of the Lactobacillus casei supragenome and its influence in species evolution and lifestyle adaptation

    Journal: BMC Genomics

    doi: 10.1186/1471-2164-13-533

    Hierarchical clustering of 17 L . casei strains based on overall gene content. Members of each cluster were aligned using Mauve, and vertical red lines indicate contig boundaries. Locally Collinear Blocks (LCBs) are colored to reveal harmonization within each cluster, but do not have identity to LCBs of the same color outside a particular cluster. The Lactobacillus casei BL23 genome was used as a reference to order and orientate contigs for strains included in all clusters except B, where strains 21/1, 32G, and Lc-10 were ordered using the L . casei Zhang genome as a reference, and strains CRF28 and 12A were ordered based on L . casei ATCC 334. The BL23 alignment is shown with cluster B to compare genomic similarity.
    Figure Legend Snippet: Hierarchical clustering of 17 L . casei strains based on overall gene content. Members of each cluster were aligned using Mauve, and vertical red lines indicate contig boundaries. Locally Collinear Blocks (LCBs) are colored to reveal harmonization within each cluster, but do not have identity to LCBs of the same color outside a particular cluster. The Lactobacillus casei BL23 genome was used as a reference to order and orientate contigs for strains included in all clusters except B, where strains 21/1, 32G, and Lc-10 were ordered using the L . casei Zhang genome as a reference, and strains CRF28 and 12A were ordered based on L . casei ATCC 334. The BL23 alignment is shown with cluster B to compare genomic similarity.

    Techniques Used:

    7) Product Images from "Characterization of a potential ABC-type bacteriocin exporter protein from Treponema denticola"

    Article Title: Characterization of a potential ABC-type bacteriocin exporter protein from Treponema denticola

    Journal: BMC Oral Health

    doi: 10.1186/s12903-016-0243-7

    Multiple sequence alignments of bacteriocin ABC transporters. Cysteine and histidine, which are part of the putative active site of the peptidase family C39B, as well as glutamine, which contributes to the oxyanion pore in other cysteine protease families, are marked with * and indicated in yellow. The ATP-binding site and ABC transporter signature motifs are indicated in yellow and marked with † and #, respectively. The alignment was carried out using the program Genetyx-Mac 16.0.9. C. divergens : ATP-dependent transporter of Carnobacterium divergens , L. lactis : Lactococcin-A transport/processing ATP-binding protein LcnC of Lactococcus lactis subsp. lactis, L. mesenteroides : Mesentericin-Y105 transport/processing ATP-binding protein MesD of Leuconostoc mesenteroides, P. acidilactici : Pediocin PA-1 transport/processing ATP-binding protein PedD of Pediococcus acidilactici, TDE_0425: tepA1 , TDE_0719: tepA2 , TDE_2431: tepA3
    Figure Legend Snippet: Multiple sequence alignments of bacteriocin ABC transporters. Cysteine and histidine, which are part of the putative active site of the peptidase family C39B, as well as glutamine, which contributes to the oxyanion pore in other cysteine protease families, are marked with * and indicated in yellow. The ATP-binding site and ABC transporter signature motifs are indicated in yellow and marked with † and #, respectively. The alignment was carried out using the program Genetyx-Mac 16.0.9. C. divergens : ATP-dependent transporter of Carnobacterium divergens , L. lactis : Lactococcin-A transport/processing ATP-binding protein LcnC of Lactococcus lactis subsp. lactis, L. mesenteroides : Mesentericin-Y105 transport/processing ATP-binding protein MesD of Leuconostoc mesenteroides, P. acidilactici : Pediocin PA-1 transport/processing ATP-binding protein PedD of Pediococcus acidilactici, TDE_0425: tepA1 , TDE_0719: tepA2 , TDE_2431: tepA3

    Techniques Used: Sequencing, Binding Assay

    Southern blot analysis of tepA1 ( a ), tepA2 ( b ) , and tepA3 ( c ). Genomic DNA from T. denticola strains was digested with Hin dIII. 1: genomic DNA from ATCC 33520, 2: genomic DNA from ATCC 33521, genomic DNA from ATCC 35404, 4: genomic DNA from ATCC 35405, 5: genomic DNA from GM1
    Figure Legend Snippet: Southern blot analysis of tepA1 ( a ), tepA2 ( b ) , and tepA3 ( c ). Genomic DNA from T. denticola strains was digested with Hin dIII. 1: genomic DNA from ATCC 33520, 2: genomic DNA from ATCC 33521, genomic DNA from ATCC 35404, 4: genomic DNA from ATCC 35405, 5: genomic DNA from GM1

    Techniques Used: Southern Blot

    Expression of tep and TDE_0820 in the presence or absence of chloramphenicol. Expression of tepA1 ( a ), tepB1 ( b ), tepA3 ( c ) , tepB3 ( d ), TDE_0820 ( e ) in the presence or absence of chloramphenicol in the wild-type and tepA2 -deficient mutant KT-3. Expression levels of each gene were normalized using 16S rRNA levels as internal controls and are expressed as a fold modulation relative to the wild-type strain grown without chloramphenicol. Experiments were performed three times in triplicate. Data are presented as the mean ± SD ( n = 9). * P
    Figure Legend Snippet: Expression of tep and TDE_0820 in the presence or absence of chloramphenicol. Expression of tepA1 ( a ), tepB1 ( b ), tepA3 ( c ) , tepB3 ( d ), TDE_0820 ( e ) in the presence or absence of chloramphenicol in the wild-type and tepA2 -deficient mutant KT-3. Expression levels of each gene were normalized using 16S rRNA levels as internal controls and are expressed as a fold modulation relative to the wild-type strain grown without chloramphenicol. Experiments were performed three times in triplicate. Data are presented as the mean ± SD ( n = 9). * P

    Techniques Used: Expressing, Mutagenesis

    8) Product Images from "Lactococcus lactis M4, a potential host for the expression of heterologous proteins"

    Article Title: Lactococcus lactis M4, a potential host for the expression of heterologous proteins

    Journal: Microbial Cell Factories

    doi: 10.1186/1475-2859-10-28

    Total DNA isolation of L. lactis subsp. lactis M4 . Lane M: λ/ Hind III marker (Fermentas); Lanes 1 to 3: L. lactis M4.
    Figure Legend Snippet: Total DNA isolation of L. lactis subsp. lactis M4 . Lane M: λ/ Hind III marker (Fermentas); Lanes 1 to 3: L. lactis M4.

    Techniques Used: DNA Extraction, Marker

    PCR-amplified partial 16S rRNA gene using P1 and P4 primers . Lane M: GeneRuler™ DNA Ladder Mix (Fermentas); Lanes 1 to 17: milk isolates; Lane 18: L. lactis subsp. cremoris MG1363; Lane 19: L. lactis subsp. lactis ATCC 11454; Lane 20: negative control.
    Figure Legend Snippet: PCR-amplified partial 16S rRNA gene using P1 and P4 primers . Lane M: GeneRuler™ DNA Ladder Mix (Fermentas); Lanes 1 to 17: milk isolates; Lane 18: L. lactis subsp. cremoris MG1363; Lane 19: L. lactis subsp. lactis ATCC 11454; Lane 20: negative control.

    Techniques Used: Polymerase Chain Reaction, Amplification, Negative Control

    9) Product Images from "Genome mining of astaxanthin biosynthetic genes from Sphingomonas sp. ATCC 55669 for heterologous overproduction in Escherichia coli"

    Article Title: Genome mining of astaxanthin biosynthetic genes from Sphingomonas sp. ATCC 55669 for heterologous overproduction in Escherichia coli

    Journal: Biotechnology Journal

    doi: 10.1002/biot.201400827

    Targeted engineering E. coli for astaxanthin overproduction. ( A ) Over‐production of the intermediates IPP and DMAPP was achieved by endogenous acetyl‐CoA via the MVA pathway. AtoB (acetyl‐CoA acetyltransferase) and Idi were from E. coli . Erg13 (3‐hydroxy‐3‐methylglutaryl‐CoA synthase), tHMG1 (a truncated version of 3‐hydroxy‐3‐methylglutaryl‐CoA reductase), ERG12 (mevalonate kinase), ERG8 (phosphomevalonate kinase), and MVD1 (mevalonate pyrophosphate decarboxylase) were from Saccharomyces cerevisiae . Plasmids pMH1 and pFZ81 were controlled by the P lac promoter. ( B ) Overproduction of astaxanthin from intermediates IPP and DMAPP via the carotenoid pathway. CrtEIB was from Pantoea ananatis ; crtYZ was from Pantoea agglomerans ; crtW was from Brevundimonas sp. SD212; idi was from E. coli ; and crtEs p1 , crtE sp2 , crtE sp3 , crtZ sp1 , and crtZ sp2 were from Sphingomonas sp. ATCC 55669. Gene expression was controlled by the P T7 promoter. ( C ) Production of astaxanthin in engineered strains from A1 to A6 detected by HPLC.
    Figure Legend Snippet: Targeted engineering E. coli for astaxanthin overproduction. ( A ) Over‐production of the intermediates IPP and DMAPP was achieved by endogenous acetyl‐CoA via the MVA pathway. AtoB (acetyl‐CoA acetyltransferase) and Idi were from E. coli . Erg13 (3‐hydroxy‐3‐methylglutaryl‐CoA synthase), tHMG1 (a truncated version of 3‐hydroxy‐3‐methylglutaryl‐CoA reductase), ERG12 (mevalonate kinase), ERG8 (phosphomevalonate kinase), and MVD1 (mevalonate pyrophosphate decarboxylase) were from Saccharomyces cerevisiae . Plasmids pMH1 and pFZ81 were controlled by the P lac promoter. ( B ) Overproduction of astaxanthin from intermediates IPP and DMAPP via the carotenoid pathway. CrtEIB was from Pantoea ananatis ; crtYZ was from Pantoea agglomerans ; crtW was from Brevundimonas sp. SD212; idi was from E. coli ; and crtEs p1 , crtE sp2 , crtE sp3 , crtZ sp1 , and crtZ sp2 were from Sphingomonas sp. ATCC 55669. Gene expression was controlled by the P T7 promoter. ( C ) Production of astaxanthin in engineered strains from A1 to A6 detected by HPLC.

    Techniques Used: Expressing, High Performance Liquid Chromatography

    The complete genome of Sphingomonas sp. ATCC 55669 contained three circular DNAs: one chromosome ( A ) and two megaplasmids ( B , C ). From innermost to outermost circles: GC content; GC‐skew ([G – C] / [G + C]); methylation information of forward sequences (red, m4C; blue, 6mA); methylation information of reverse sequences (pink, m4C; cyan, 6mA); predicted genes color‐coded by COG categories of forward and reverse strands, respectively; and genes related to the non‐mevalonate pathway and carotenoid biosynthesis. All genes putatively involved in astaxanthin biosynthesis “ (gene names noted in the outer circle) showed a discrete distribution throughout the chromosome.
    Figure Legend Snippet: The complete genome of Sphingomonas sp. ATCC 55669 contained three circular DNAs: one chromosome ( A ) and two megaplasmids ( B , C ). From innermost to outermost circles: GC content; GC‐skew ([G – C] / [G + C]); methylation information of forward sequences (red, m4C; blue, 6mA); methylation information of reverse sequences (pink, m4C; cyan, 6mA); predicted genes color‐coded by COG categories of forward and reverse strands, respectively; and genes related to the non‐mevalonate pathway and carotenoid biosynthesis. All genes putatively involved in astaxanthin biosynthesis “ (gene names noted in the outer circle) showed a discrete distribution throughout the chromosome.

    Techniques Used: Methylation

    Proposed astaxanthin biosynthetic pathway in Sphingomonas sp. ATCC 55669. ( A ) In the strain Sphingomonas sp. ATCC 55669, upstream events via the MEP pathway to get the precursors IPP and DMAPP. ( B ) The downstream carotenoid pathway involved the conversion of intermediates IPP and DMAPP to astaxanthin. DXS, 1‐deoxy‐D‐xylulose‐5‐phosphate synthase; DXR, 1‐deoxy‐D‐xylulose‐5‐phosphate reductoisomerase; IspD, 2‐ C ‐methyl‐D‐erythritol 4‐phosphate cytidylyltransferase; IspE, 4‐diphosphocytidyl‐2‐ C ‐methyl‐D‐erythritol kinase; IspF, 2‐ C ‐methyl‐D‐erythritol 2,4‐cyclodiphosphate synthase; IspG, (E)‐4‐hydroxy‐3‐methylbut‐2‐enyl‐diphosphate synthase; IspH, 4‐hydroxy‐3‐methylbut‐2‐enyl diphosphate reductase. G‐3‐P,D‐glyceraldehyde 3‐phosphate; DXP, 1‐deoxy‐D‐xylulose 5‐phosphate; CDP‐ME, 4‐(cytidine 5'‐diphospho)‐2‐ C ‐methyl‐D‐erythritol; CDP‐ME2P, 2‐phospho‐4‐(cytidine 5'‐diphospho)‐2‐ C ‐methyl‐D‐erythritol; ME‐cPP, 2‐ C ‐methyl‐D‐erythritol 2,4‐cyclodiphosphate; HMBPP, 1‐hydroxy‐2‐methyl‐2‐butenyl 4‐diphosphate.
    Figure Legend Snippet: Proposed astaxanthin biosynthetic pathway in Sphingomonas sp. ATCC 55669. ( A ) In the strain Sphingomonas sp. ATCC 55669, upstream events via the MEP pathway to get the precursors IPP and DMAPP. ( B ) The downstream carotenoid pathway involved the conversion of intermediates IPP and DMAPP to astaxanthin. DXS, 1‐deoxy‐D‐xylulose‐5‐phosphate synthase; DXR, 1‐deoxy‐D‐xylulose‐5‐phosphate reductoisomerase; IspD, 2‐ C ‐methyl‐D‐erythritol 4‐phosphate cytidylyltransferase; IspE, 4‐diphosphocytidyl‐2‐ C ‐methyl‐D‐erythritol kinase; IspF, 2‐ C ‐methyl‐D‐erythritol 2,4‐cyclodiphosphate synthase; IspG, (E)‐4‐hydroxy‐3‐methylbut‐2‐enyl‐diphosphate synthase; IspH, 4‐hydroxy‐3‐methylbut‐2‐enyl diphosphate reductase. G‐3‐P,D‐glyceraldehyde 3‐phosphate; DXP, 1‐deoxy‐D‐xylulose 5‐phosphate; CDP‐ME, 4‐(cytidine 5'‐diphospho)‐2‐ C ‐methyl‐D‐erythritol; CDP‐ME2P, 2‐phospho‐4‐(cytidine 5'‐diphospho)‐2‐ C ‐methyl‐D‐erythritol; ME‐cPP, 2‐ C ‐methyl‐D‐erythritol 2,4‐cyclodiphosphate; HMBPP, 1‐hydroxy‐2‐methyl‐2‐butenyl 4‐diphosphate.

    Techniques Used: Conditioned Place Preference

    10) Product Images from "Preclinical evaluation of bacterially produced RSV-G protein vaccine: Strong protection against RSV challenge in cotton rat model"

    Article Title: Preclinical evaluation of bacterially produced RSV-G protein vaccine: Strong protection against RSV challenge in cotton rat model

    Journal: Scientific Reports

    doi: 10.1038/srep42428

    Neutralizing antibody response following RSV-G protein, FI-RSV and live RSV experimental infection. ( A ) Schematic representation of cotton rat immunization and challenge schedule. Inbred female  Sigmodon hispidus  cotton rats between 6 and 8 weeks of age were immunized i.m. with PBS (Gps A-B), 5 μg of unadjuvanted or Emulsigen-adjuvanted REG ( R ecombinant  E . coli  produced  G ) (Gps C-D), or with FI-RSV (Gp E) on days 0 and 28 in groups A thru E, or were infected intranasally (i.n.) with 0.1 ml of RSV/A2 at 10 5  pfu per rat (Gp F). Blood was collected by eye-bleed on days 0, 28 and 49. On day 49, animals were either mock challenge intranasally with 0.1 ml of PBS (Gp A), or with 0.1 ml of RSV-A2 virus at 10 5  pfu per animal (10 animals per group) (Gps B-F). Cotton rats were sacrificed on days 2 or 5 post-challenge wherein lungs and nose tissues were collected. ( B ) Sera from individual cotton rats collected at pre-vaccination (day 0) and 3 weeks post second immunization (day 49) were tested for neutralization in a plaque reduction neutralization test (PRNT) against the homologous RSV-A2 strain and heterologous RSV-B1 strain. Neutralizing antibody titers represent 50% inhibition of plaque numbers. Statistical significance was tested with one-way ANOVA and Bonferroni multiple comparisons tests. ***p 
    Figure Legend Snippet: Neutralizing antibody response following RSV-G protein, FI-RSV and live RSV experimental infection. ( A ) Schematic representation of cotton rat immunization and challenge schedule. Inbred female Sigmodon hispidus cotton rats between 6 and 8 weeks of age were immunized i.m. with PBS (Gps A-B), 5 μg of unadjuvanted or Emulsigen-adjuvanted REG ( R ecombinant E . coli produced G ) (Gps C-D), or with FI-RSV (Gp E) on days 0 and 28 in groups A thru E, or were infected intranasally (i.n.) with 0.1 ml of RSV/A2 at 10 5 pfu per rat (Gp F). Blood was collected by eye-bleed on days 0, 28 and 49. On day 49, animals were either mock challenge intranasally with 0.1 ml of PBS (Gp A), or with 0.1 ml of RSV-A2 virus at 10 5 pfu per animal (10 animals per group) (Gps B-F). Cotton rats were sacrificed on days 2 or 5 post-challenge wherein lungs and nose tissues were collected. ( B ) Sera from individual cotton rats collected at pre-vaccination (day 0) and 3 weeks post second immunization (day 49) were tested for neutralization in a plaque reduction neutralization test (PRNT) against the homologous RSV-A2 strain and heterologous RSV-B1 strain. Neutralizing antibody titers represent 50% inhibition of plaque numbers. Statistical significance was tested with one-way ANOVA and Bonferroni multiple comparisons tests. ***p 

    Techniques Used: Infection, Produced, Neutralization, Plaque Reduction Neutralization Test, Inhibition

    11) Product Images from "Broad-Spectrum Inhibition of Respiratory Virus Infection by MicroRNA Mimics Targeting p38 MAPK Signaling"

    Article Title: Broad-Spectrum Inhibition of Respiratory Virus Infection by MicroRNA Mimics Targeting p38 MAPK Signaling

    Journal: Molecular Therapy. Nucleic Acids

    doi: 10.1016/j.omtn.2017.03.008

    Reductions in IAV and RSV Titer following Treatments with miRNA Mimics A549 cells were transfected with 25 nM miRNA mimics or controls. A direct-targeting viral siRNA (siIAV or siRSV) served as a positive antiviral control (gray bar), while negative controls were as follows: C. elegans miRNA mimic 1 (C.elegans 1), RISC-free siRNA, Lipofectamine, and virus infection alone (black bars). (A–E) After 48-hr transfection, media was removed and cells were infected with (A) IAV WSN H1N1 at MOI 0.1 for 24 hr, (B) IAV PR8 H1N1 at MOI 0.1 for 12 hr, (C) IAV Udorn H3N2 at MOI 0.1 for 24 hr, (D) RSV-A2 at MOI 0.01 for 72 hr, and (E) RSV-BT2a at MOI 0.1 for 72 hr, when the supernatant was removed and assayed for virus. (F) Uninfected cells were analyzed for cell viability at 48 hr post-transfection (n = 12). The results are displayed to show the most potent antiviral mimics (from left to right along the x axis), with solid lines denoting 0% inhibition, dotted lines denoting the 75% activity cutoff, and blue bars highlighting miRNAs chosen for further analysis. Viral titer results for each virus are shown as the mean ± SEM of n = 6. Significant differences between virus control and miRNA mimics and siRNAs are indicated (*p
    Figure Legend Snippet: Reductions in IAV and RSV Titer following Treatments with miRNA Mimics A549 cells were transfected with 25 nM miRNA mimics or controls. A direct-targeting viral siRNA (siIAV or siRSV) served as a positive antiviral control (gray bar), while negative controls were as follows: C. elegans miRNA mimic 1 (C.elegans 1), RISC-free siRNA, Lipofectamine, and virus infection alone (black bars). (A–E) After 48-hr transfection, media was removed and cells were infected with (A) IAV WSN H1N1 at MOI 0.1 for 24 hr, (B) IAV PR8 H1N1 at MOI 0.1 for 12 hr, (C) IAV Udorn H3N2 at MOI 0.1 for 24 hr, (D) RSV-A2 at MOI 0.01 for 72 hr, and (E) RSV-BT2a at MOI 0.1 for 72 hr, when the supernatant was removed and assayed for virus. (F) Uninfected cells were analyzed for cell viability at 48 hr post-transfection (n = 12). The results are displayed to show the most potent antiviral mimics (from left to right along the x axis), with solid lines denoting 0% inhibition, dotted lines denoting the 75% activity cutoff, and blue bars highlighting miRNAs chosen for further analysis. Viral titer results for each virus are shown as the mean ± SEM of n = 6. Significant differences between virus control and miRNA mimics and siRNAs are indicated (*p

    Techniques Used: Transfection, Infection, Inhibition, Activity Assay

    12) Product Images from "Genetic dissection of the biosynthetic route to gentamicin A2 by heterologous expression of its minimal gene set"

    Article Title: Genetic dissection of the biosynthetic route to gentamicin A2 by heterologous expression of its minimal gene set

    Journal:

    doi: 10.1073/pnas.0803164105

    GtmE Acts as an UDP-Xylose Glycosyltransferase to Generate Gentamicin A2 (4a).
    Figure Legend Snippet: GtmE Acts as an UDP-Xylose Glycosyltransferase to Generate Gentamicin A2 (4a).

    Techniques Used:

    HPLC-ESI-MS/MS chromatograms for the glycosyltransferase-catalyzed production of 2′- N -acetylparomamine ( 2a ) and gentamicin A2 ( 4a ). ( A–C ) GtmG assay using cell-free extracts from S. venezuelae strain harboring pYJ498 ( A ) without exogenous
    Figure Legend Snippet: HPLC-ESI-MS/MS chromatograms for the glycosyltransferase-catalyzed production of 2′- N -acetylparomamine ( 2a ) and gentamicin A2 ( 4a ). ( A–C ) GtmG assay using cell-free extracts from S. venezuelae strain harboring pYJ498 ( A ) without exogenous

    Techniques Used: High Performance Liquid Chromatography, Mass Spectrometry

    GtmE Acts as an UDP-Xylose Glycosyltransferase to Generate Gentamicin A2 (4a).
    Figure Legend Snippet: GtmE Acts as an UDP-Xylose Glycosyltransferase to Generate Gentamicin A2 (4a).

    Techniques Used:

    13) Product Images from "Differentiation of Clostridium botulinum Serotype A Strains by Multiple-Locus Variable-Number Tandem-Repeat Analysis ▿ Serotype A Strains by Multiple-Locus Variable-Number Tandem-Repeat Analysis ▿ †"

    Article Title: Differentiation of Clostridium botulinum Serotype A Strains by Multiple-Locus Variable-Number Tandem-Repeat Analysis ▿ Serotype A Strains by Multiple-Locus Variable-Number Tandem-Repeat Analysis ▿ †

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.01539-07

    VNTR profiles within four C. botulinum strains. Four of the 10 VNTR profiles were overlaid to illustrate the different fragment sizes generated in four strains. VNTR 1, 3, 4, and 7 fragment sizes (in base pairs) are shown in blue for the BoNT/A1 strains
    Figure Legend Snippet: VNTR profiles within four C. botulinum strains. Four of the 10 VNTR profiles were overlaid to illustrate the different fragment sizes generated in four strains. VNTR 1, 3, 4, and 7 fragment sizes (in base pairs) are shown in blue for the BoNT/A1 strains

    Techniques Used: Generated

    MLVA of 61 C. botulinum strains. Fifty-nine BoNT/A1-A4-producing strains and three bivalent B strains were differentiated into 38 genotypes by use of 10 VNTR loci. The completed genomic sequences of BoNT/A1 strains ATCC 3502, ATCC 19397, and A150 were
    Figure Legend Snippet: MLVA of 61 C. botulinum strains. Fifty-nine BoNT/A1-A4-producing strains and three bivalent B strains were differentiated into 38 genotypes by use of 10 VNTR loci. The completed genomic sequences of BoNT/A1 strains ATCC 3502, ATCC 19397, and A150 were

    Techniques Used: Genomic Sequencing

    14) Product Images from "Enhanced Antitumor Activity of Murine-Human Hybrid T-Cell Receptor (TCR) in Human Lymphocytes Is Associated with Improved Pairing and TCR/CD3 Stability"

    Article Title: Enhanced Antitumor Activity of Murine-Human Hybrid T-Cell Receptor (TCR) in Human Lymphocytes Is Associated with Improved Pairing and TCR/CD3 Stability

    Journal:

    doi: 10.1158/0008-5472.CAN-06-1450

    A, TCR competition assay. TCR-deficient Jurkat RT3-T3.5 cells were electroporated with 1 μg of each chain of the MART-HH ( white columns ) or MART-HM ( gray columns ) along with 1 μg of each chain of the competitor TCR [for p53-MH, we also
    Figure Legend Snippet: A, TCR competition assay. TCR-deficient Jurkat RT3-T3.5 cells were electroporated with 1 μg of each chain of the MART-HH ( white columns ) or MART-HM ( gray columns ) along with 1 μg of each chain of the competitor TCR [for p53-MH, we also

    Techniques Used: Competitive Binding Assay

    15) Product Images from "Prevalence of the sodC Gene in Nontypeable Haemophilus influenzae and Haemophilus haemolyticus by Microarray-Based Hybridization ▿"

    Article Title: Prevalence of the sodC Gene in Nontypeable Haemophilus influenzae and Haemophilus haemolyticus by Microarray-Based Hybridization ▿

    Journal: Journal of Clinical Microbiology

    doi: 10.1128/JCM.01416-09

    Southern hybridization of a sodC gene probe to microarray-positive NT H. influenzae strains. Lane 1, negative-control H. influenzae strain Rd; lane 2, positive-control H. haemolyticus strain 65P28H9; lanes 3 to 10, eight representative H. influenzae strains, P18-25, P05-24, O05-21, I8809, C21-21, O14-21, C14-21, and C09-23, respectively.
    Figure Legend Snippet: Southern hybridization of a sodC gene probe to microarray-positive NT H. influenzae strains. Lane 1, negative-control H. influenzae strain Rd; lane 2, positive-control H. haemolyticus strain 65P28H9; lanes 3 to 10, eight representative H. influenzae strains, P18-25, P05-24, O05-21, I8809, C21-21, O14-21, C14-21, and C09-23, respectively.

    Techniques Used: Hybridization, Microarray, Negative Control, Positive Control

    16) Product Images from "Human Respiratory Syncytial Virus Nucleoprotein and Inclusion Bodies Antagonize the Innate Immune Response Mediated by MDA5 and MAVS"

    Article Title: Human Respiratory Syncytial Virus Nucleoprotein and Inclusion Bodies Antagonize the Innate Immune Response Mediated by MDA5 and MAVS

    Journal: Journal of Virology

    doi: 10.1128/JVI.00215-12

    hRSV IBs colocalize with MAVS and RIG-I. A549 cells were infected with RSV strain A2 at an MOI of 1.0 and incubated for 6, 12, or 24 h or were mock infected. hRSV N, MAVS, and RIG-I were detected by immunofluorescence. Column 1, RSV N staining; column
    Figure Legend Snippet: hRSV IBs colocalize with MAVS and RIG-I. A549 cells were infected with RSV strain A2 at an MOI of 1.0 and incubated for 6, 12, or 24 h or were mock infected. hRSV N, MAVS, and RIG-I were detected by immunofluorescence. Column 1, RSV N staining; column

    Techniques Used: Infection, Incubation, Immunofluorescence, Staining

    hRSV IBs colocalize with MDA5 and RIG-I. A549 cells were infected with hRSV strain A2 at an MOI of 1.0 and incubated for 6, 12, or 24 h or were mock infected. hRSV N, MDA5, and RIG-I were detected by immunofluorescence. Column 1, hRSV staining; column
    Figure Legend Snippet: hRSV IBs colocalize with MDA5 and RIG-I. A549 cells were infected with hRSV strain A2 at an MOI of 1.0 and incubated for 6, 12, or 24 h or were mock infected. hRSV N, MDA5, and RIG-I were detected by immunofluorescence. Column 1, hRSV staining; column

    Techniques Used: Infection, Incubation, Immunofluorescence, Staining

    hRSV genomic RNA colocalized with small but not large inclusion bodies (IBs). HEp-2 cells were infected with hRSV strain A2 at an MOI of 1.0 for 6, 12, or 24 h, and subsequently hRSV N was detected using immunostaining and genomic RNA detected with a
    Figure Legend Snippet: hRSV genomic RNA colocalized with small but not large inclusion bodies (IBs). HEp-2 cells were infected with hRSV strain A2 at an MOI of 1.0 for 6, 12, or 24 h, and subsequently hRSV N was detected using immunostaining and genomic RNA detected with a

    Techniques Used: Infection, Immunostaining

    PLA gave no signal in primary antibody control cells infected with RSV. Vero cells were infected with hRSV strain A2 at an MOI of 1.0 and incubated for 6 or 24 h. Cells were stained for RSV P (green) and were assayed for the interaction between RSV N
    Figure Legend Snippet: PLA gave no signal in primary antibody control cells infected with RSV. Vero cells were infected with hRSV strain A2 at an MOI of 1.0 and incubated for 6 or 24 h. Cells were stained for RSV P (green) and were assayed for the interaction between RSV N

    Techniques Used: Proximity Ligation Assay, Infection, Incubation, Staining

    hRSV N interacts with MDA5 and MAVS. Vero cells were infected with hRSV strain A2 at an MOI of 1.0 and incubated for 6 or 24 h. Cells were stained for hRSV P and were assayed for the interaction between hRSV N and either MDA5 or MAVS by PLA. Column 1,
    Figure Legend Snippet: hRSV N interacts with MDA5 and MAVS. Vero cells were infected with hRSV strain A2 at an MOI of 1.0 and incubated for 6 or 24 h. Cells were stained for hRSV P and were assayed for the interaction between hRSV N and either MDA5 or MAVS by PLA. Column 1,

    Techniques Used: Infection, Incubation, Staining, Proximity Ligation Assay

    MDA5 colocalized with isolated viral filaments. HEp-2 cells were infected with hRSV strain A2 at an MOI of 0.1, and viral filaments were isolated 4 dpi. Viral filaments were isolated by filtration and adsorbed onto cover glass for immunostaining. Column
    Figure Legend Snippet: MDA5 colocalized with isolated viral filaments. HEp-2 cells were infected with hRSV strain A2 at an MOI of 0.1, and viral filaments were isolated 4 dpi. Viral filaments were isolated by filtration and adsorbed onto cover glass for immunostaining. Column

    Techniques Used: Isolation, Infection, Filtration, Immunostaining

    Secondary antibodies do not cross-react with the primary antibodies, nor do the fluorophors bleed through to adjacent fluorescent channels. A549 cells were infected with hRSV strain A2 at an MOI of 1.0 for 24 h and were subjected to immunostaining using
    Figure Legend Snippet: Secondary antibodies do not cross-react with the primary antibodies, nor do the fluorophors bleed through to adjacent fluorescent channels. A549 cells were infected with hRSV strain A2 at an MOI of 1.0 for 24 h and were subjected to immunostaining using

    Techniques Used: Infection, Immunostaining

    17) Product Images from "Horizontal Transfer of the OXA-24 Carbapenemase Gene via Outer Membrane Vesicles: a New Mechanism of Dissemination of Carbapenem Resistance Genes in Acinetobacter baumannii ▿"

    Article Title: Horizontal Transfer of the OXA-24 Carbapenemase Gene via Outer Membrane Vesicles: a New Mechanism of Dissemination of Carbapenem Resistance Genes in Acinetobacter baumannii ▿

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00929-10

    PCR fingerprinting profile of plasmids pMMA2 and pMCU3, determined by using a set of oligonucleotides. Lane 1, PCR fingerprint of plasmids isolated from strain AbH12O-A2; lane 2, PCR fingerprint of plasmids isolated from strain ATCC 17978 transformed
    Figure Legend Snippet: PCR fingerprinting profile of plasmids pMMA2 and pMCU3, determined by using a set of oligonucleotides. Lane 1, PCR fingerprint of plasmids isolated from strain AbH12O-A2; lane 2, PCR fingerprint of plasmids isolated from strain ATCC 17978 transformed

    Techniques Used: Polymerase Chain Reaction, Isolation, Transformation Assay

    REP-PCR profile of the A. baumannii strains used in this study. Blot A, AbH12O-A2; blot B, AbH12O-CU3; blot C, ATCC 17978; blot D, ATCC 17978 transformed with OMVs from AbH12O-A2; blot E, ATCC 17978 transformed with OMVs from AbH12O-CU3.
    Figure Legend Snippet: REP-PCR profile of the A. baumannii strains used in this study. Blot A, AbH12O-A2; blot B, AbH12O-CU3; blot C, ATCC 17978; blot D, ATCC 17978 transformed with OMVs from AbH12O-A2; blot E, ATCC 17978 transformed with OMVs from AbH12O-CU3.

    Techniques Used: Polymerase Chain Reaction, Transformation Assay

    Electron microscopy micrograph of OMVs released by A. baumannii clinical strain AbH12O-A2. Vesicles were purified from broth cultures by ultracentrifugation and filtered through a 0.22-μm filter. The average diameter of the vesicles was 40 nm.
    Figure Legend Snippet: Electron microscopy micrograph of OMVs released by A. baumannii clinical strain AbH12O-A2. Vesicles were purified from broth cultures by ultracentrifugation and filtered through a 0.22-μm filter. The average diameter of the vesicles was 40 nm.

    Techniques Used: Electron Microscopy, Purification

    (A) Dose-response experiment. Shown is a dot blot analysis for detecting the presence of the bla OXA- 24 gene in OMVs released from A. baumannii clinical isolate AbH12O-A2. Blot A, 10 μl of purified plasmid pMMA2 as a positive control; blot B, 10
    Figure Legend Snippet: (A) Dose-response experiment. Shown is a dot blot analysis for detecting the presence of the bla OXA- 24 gene in OMVs released from A. baumannii clinical isolate AbH12O-A2. Blot A, 10 μl of purified plasmid pMMA2 as a positive control; blot B, 10

    Techniques Used: Dot Blot, Purification, Plasmid Preparation, Positive Control

    18) Product Images from "Comparative Proteomic Analysis of Extracellular Proteins of Clostridium perfringens Type A and Type C Strains ▿ Type A and Type C Strains ▿ †"

    Article Title: Comparative Proteomic Analysis of Extracellular Proteins of Clostridium perfringens Type A and Type C Strains ▿ Type A and Type C Strains ▿ †

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00374-10

    Relative abundances of identified exoproteins from the C. perfringens ATCC 13124 (spot numbers beginning with A) and pooled type C (spot numbers beginning with C) exoproteomes. The abundance is relative to the level of the most abundant protein on the
    Figure Legend Snippet: Relative abundances of identified exoproteins from the C. perfringens ATCC 13124 (spot numbers beginning with A) and pooled type C (spot numbers beginning with C) exoproteomes. The abundance is relative to the level of the most abundant protein on the

    Techniques Used:

    19) Product Images from "Intratumoral IL-12 Gene Therapy Results in the Crosspriming of Tc1 Cells Reactive Against Tumor-associated Stromal Antigens"

    Article Title: Intratumoral IL-12 Gene Therapy Results in the Crosspriming of Tc1 Cells Reactive Against Tumor-associated Stromal Antigens

    Journal: Molecular Therapy

    doi: 10.1038/mt.2010.295

    Splenic CD8 + T cells from HHD mice effectively treated with DC.IL12 gene therapy develop HLA-A2-restricted responses against melanoma-associated antigens . HHD mice bearing day 7 HLA-A2 neg (MART-1 + , gp100 + ) B16 melanomas were left
    Figure Legend Snippet: Splenic CD8 + T cells from HHD mice effectively treated with DC.IL12 gene therapy develop HLA-A2-restricted responses against melanoma-associated antigens . HHD mice bearing day 7 HLA-A2 neg (MART-1 + , gp100 + ) B16 melanomas were left

    Techniques Used: Mouse Assay

    20) Product Images from "Genomic Epidemiology of Clostridium botulinum Isolates from Temporally Related Cases of Infant Botulism in New South Wales, Australia"

    Article Title: Genomic Epidemiology of Clostridium botulinum Isolates from Temporally Related Cases of Infant Botulism in New South Wales, Australia

    Journal: Journal of Clinical Microbiology

    doi: 10.1128/JCM.00143-15

    bont gene phylogenetic analyses. (A) Dendrogram of selected bont/ A nucleotide sequences. The bont/ A sequences from NSW1_A2, NSW2_A2, and NSW3_A2 clustered with bont /A2 gene sequences. (B) Dendrogram of selected bont/ B genes, showing the NSW4_B6 gene clustered
    Figure Legend Snippet: bont gene phylogenetic analyses. (A) Dendrogram of selected bont/ A nucleotide sequences. The bont/ A sequences from NSW1_A2, NSW2_A2, and NSW3_A2 clustered with bont /A2 gene sequences. (B) Dendrogram of selected bont/ B genes, showing the NSW4_B6 gene clustered

    Techniques Used:

    21) Product Images from "Cytotoxic Indolocarbazoles from Actinomadura melliaura ATCC 39691"

    Article Title: Cytotoxic Indolocarbazoles from Actinomadura melliaura ATCC 39691

    Journal: Journal of natural products

    doi: 10.1021/acs.jnatprod.5b00429

    Key NOESY correlations for indolocarbazoles 1 , 2 , and 4 .
    Figure Legend Snippet: Key NOESY correlations for indolocarbazoles 1 , 2 , and 4 .

    Techniques Used:

    1 H– 1 H–COSY and selected HMBC correlations for indolocarbazoles 1 – 4 .
    Figure Legend Snippet: 1 H– 1 H–COSY and selected HMBC correlations for indolocarbazoles 1 – 4 .

    Techniques Used:

    22) Product Images from "Identification and Evaluation of a Highly Effective Fusion Inhibitor for Human Metapneumovirus ▿"

    Article Title: Identification and Evaluation of a Highly Effective Fusion Inhibitor for Human Metapneumovirus ▿

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00793-07

    Alignment of the HRA and HRB amino acid sequences from hMPV groups A1, A2, B1, and B2, as well as from hRSV (strain A2) and hPIV-3 fusion proteins. The sequences were aligned using ClustalW with BioEdit software (version 7.0.5.2). (A) Alignment of hMPV, hRSV, and hPIV-3 HRA sequences. Gray letters are amino acids added to the HRA sequence to form the HRA2 peptide. (B) Alignment of hMPV, hRSV, and hPIV-3 HRB sequences. (C) Amino acid identity for HRA, HRA2, and HRB sequences of hMPV, hRSV, and hPIV-3 fusion protein sequences. hMPV clinical strains C-85473 and NL-001 (GenBank accession no. AF371337) were used as representatives of the hMPV A1 group; hMPV Can97-83 (GenBank AY145296) was used as representative of the hMPV A2 group; hMPV Can97-82 (GenBank AY145295) was used as representative of the hMPV B1 group; hMPV Can98-75 (GenBank AY145289) was used as representative of the hMPV B2 group; hRSV strain A2 (GenBank M11486) and hPIV-3 (GenBank M14892) were also used in this alignment.
    Figure Legend Snippet: Alignment of the HRA and HRB amino acid sequences from hMPV groups A1, A2, B1, and B2, as well as from hRSV (strain A2) and hPIV-3 fusion proteins. The sequences were aligned using ClustalW with BioEdit software (version 7.0.5.2). (A) Alignment of hMPV, hRSV, and hPIV-3 HRA sequences. Gray letters are amino acids added to the HRA sequence to form the HRA2 peptide. (B) Alignment of hMPV, hRSV, and hPIV-3 HRB sequences. (C) Amino acid identity for HRA, HRA2, and HRB sequences of hMPV, hRSV, and hPIV-3 fusion protein sequences. hMPV clinical strains C-85473 and NL-001 (GenBank accession no. AF371337) were used as representatives of the hMPV A1 group; hMPV Can97-83 (GenBank AY145296) was used as representative of the hMPV A2 group; hMPV Can97-82 (GenBank AY145295) was used as representative of the hMPV B1 group; hMPV Can98-75 (GenBank AY145289) was used as representative of the hMPV B2 group; hRSV strain A2 (GenBank M11486) and hPIV-3 (GenBank M14892) were also used in this alignment.

    Techniques Used: Software, Sequencing

    23) Product Images from "The Transcriptional Cycle Is Suited to Daytime N2 Fixation in the Unicellular Cyanobacterium “Candidatus Atelocyanobacterium thalassa” (UCYN-A)"

    Article Title: The Transcriptional Cycle Is Suited to Daytime N2 Fixation in the Unicellular Cyanobacterium “Candidatus Atelocyanobacterium thalassa” (UCYN-A)

    Journal: mBio

    doi: 10.1128/mBio.02495-18

    Transcription of genes for replication and cell division in UCYN-A. (A) (Upper panel) Diel transcription patterns for cell division and replication genes in UCYN-A1 and UCYN-A2 over the light-dark cycle. Hierarchical clustering of genes was based on Pearson correlations between their transcription profiles. The transcription values of the genes were standardized at each time point, and the blue-red scale shows by how many standard deviations a transcription value was lower (blue) or higher (red) than the mean transcription values over the diel cycle (Z score). The Gene ID and the gene product corresponding to each gene for UCYN-A1 and UCYN-A2 are shown. Time periods are indicated on the x ). (B) Two big clusters of UCYN-A2 cells and the attached haptophyte host. (Left panel) The nucleus of the host and the UCYN-A2 cells were visualized with DAPI stain (blue). (Right panel) The UCYN-A2 strain (red) and its haptophyte host (green). (C) Two different associations of UCYN-A2 with its haptophyte dividing in samples from Scripps Pier.
    Figure Legend Snippet: Transcription of genes for replication and cell division in UCYN-A. (A) (Upper panel) Diel transcription patterns for cell division and replication genes in UCYN-A1 and UCYN-A2 over the light-dark cycle. Hierarchical clustering of genes was based on Pearson correlations between their transcription profiles. The transcription values of the genes were standardized at each time point, and the blue-red scale shows by how many standard deviations a transcription value was lower (blue) or higher (red) than the mean transcription values over the diel cycle (Z score). The Gene ID and the gene product corresponding to each gene for UCYN-A1 and UCYN-A2 are shown. Time periods are indicated on the x ). (B) Two big clusters of UCYN-A2 cells and the attached haptophyte host. (Left panel) The nucleus of the host and the UCYN-A2 cells were visualized with DAPI stain (blue). (Right panel) The UCYN-A2 strain (red) and its haptophyte host (green). (C) Two different associations of UCYN-A2 with its haptophyte dividing in samples from Scripps Pier.

    Techniques Used: Staining

    Schematic model of UCYN-A showing the possible main cellular functions, metabolic pathways, and transporters.
    Figure Legend Snippet: Schematic model of UCYN-A showing the possible main cellular functions, metabolic pathways, and transporters.

    Techniques Used:

    Network showing the Pearson correlation for gene transcriptions in the unicellular N 2 -fixing cyanobacteria Cyanothece sp. ATCC 51142 ( Cyanothece ), C. watsonii WH 8501 ( Crocosphaera ), and UCYN-A. Shown here are key genes in major metabolic pathways with distinct diel transcription patterns. The genes are shown as connected if their correlation coefficient for transcription patterns is higher than 0.2. PPP, pentose phosphate pathway; PSI, photosystem I.
    Figure Legend Snippet: Network showing the Pearson correlation for gene transcriptions in the unicellular N 2 -fixing cyanobacteria Cyanothece sp. ATCC 51142 ( Cyanothece ), C. watsonii WH 8501 ( Crocosphaera ), and UCYN-A. Shown here are key genes in major metabolic pathways with distinct diel transcription patterns. The genes are shown as connected if their correlation coefficient for transcription patterns is higher than 0.2. PPP, pentose phosphate pathway; PSI, photosystem I.

    Techniques Used:

    24) Product Images from "Effect of Previous Respiratory Syncytial Virus Infection on Murine Immune Responses to F and G Protein-Containing Virus-Like Particles"

    Article Title: Effect of Previous Respiratory Syncytial Virus Infection on Murine Immune Responses to F and G Protein-Containing Virus-Like Particles

    Journal: Journal of Virology

    doi: 10.1128/JVI.00087-19

    Neutralizing antibody titers. Neutralizing antibody titers with time after boosts of RSV-primed (set 1) and VLP-primed (set 2) animals are shown in panels A and B, respectively. Aliquots of sera from each group of animals acquired at each time point after the VLP or RSV boost were pooled. The experiment was accomplished twice for the RSV-primed animals (10 animals/group) and three times for the VLP-primed animals (15 animals/group). Serum from each experiment at each time point was pooled separately and titers of each pool determined separately. The NA titers are the reciprocal of the dilution of sera resulting in a 50% reduction of virus titer in a plaque reduction assay. The results are averages from 4 to 6 separate determinations on each pool. Since the results from each of the pools from equivalent groups were not statistically different, the results were combined. The averages and standard deviations are shown. Note the change in y axis scales to visualize lower titers. (C) Titers in each group of animals in the two sets of mice at week 4 after the VLP boost with statistically significant differences in values for between groups within a set and between groups in the two sets of animals. The significance of results at 17 weeks postboost is indicated in panel A. There were no significant differences at late times in VLP-primed animals. (D) RSV, strain B, neutralization titers using sera from animals primed with either RSV or with pre-F VLPs, boosted with pre-F VLPs, and harvested at 4 weeks postboost. Results are from pools of sera from two separate experiments, and the assay with each pool was accomplished twice. *, P = 0.05; **, P = 0.005; ***, P = 0.0005.
    Figure Legend Snippet: Neutralizing antibody titers. Neutralizing antibody titers with time after boosts of RSV-primed (set 1) and VLP-primed (set 2) animals are shown in panels A and B, respectively. Aliquots of sera from each group of animals acquired at each time point after the VLP or RSV boost were pooled. The experiment was accomplished twice for the RSV-primed animals (10 animals/group) and three times for the VLP-primed animals (15 animals/group). Serum from each experiment at each time point was pooled separately and titers of each pool determined separately. The NA titers are the reciprocal of the dilution of sera resulting in a 50% reduction of virus titer in a plaque reduction assay. The results are averages from 4 to 6 separate determinations on each pool. Since the results from each of the pools from equivalent groups were not statistically different, the results were combined. The averages and standard deviations are shown. Note the change in y axis scales to visualize lower titers. (C) Titers in each group of animals in the two sets of mice at week 4 after the VLP boost with statistically significant differences in values for between groups within a set and between groups in the two sets of animals. The significance of results at 17 weeks postboost is indicated in panel A. There were no significant differences at late times in VLP-primed animals. (D) RSV, strain B, neutralization titers using sera from animals primed with either RSV or with pre-F VLPs, boosted with pre-F VLPs, and harvested at 4 weeks postboost. Results are from pools of sera from two separate experiments, and the assay with each pool was accomplished twice. *, P = 0.05; **, P = 0.005; ***, P = 0.0005.

    Techniques Used: Mouse Assay, Neutralization

    Titers of anti-F protein IgG. (A and B) The level (ng/ml) of anti-F protein antibodies, determined by ELISA in pooled sera, that binds to the soluble prefusion or postfusion F protein, with time after VLP boosts of RSV-primed animals or a second RSV infection. The values shown are averages with standard deviations from 4 to 5 separate determinations on each of two pools of sera from two different experiments. Values obtained with the two pools of sera from each time point were not statistically significantly different and therefore were combined. (C and D) Levels (ng/ml) of pre-F binding or post-F binding antibodies with time in pools of sera from VLP-primed, VLP-boosted animals. The values shown are averages, with standard deviations, from 4 to 5 separate determinations on each of three pooled sera from three different experiments. Values obtained with each of the three pools of sera were not statistically significantly different and therefore were combined. (E and F) Level (ng/ml) of pre-F (E) or post-F (F) binding antibodies at 4 weeks postboost (the time of maximal NA titers) are shown for clear presentation of statistically significant differences between groups and between the sets of mice. (G and H) Statistically significant differences, at 4 weeks postboost, between pre-F and post-F protein binding antibodies within each set of animals. There were no significant differences between VLP-primed animals at late times postboost. *, P = 0.05; **, P = 0.005; ***, P = 0.0005; ****, P = 0.00005. Only significant differences are indicated.
    Figure Legend Snippet: Titers of anti-F protein IgG. (A and B) The level (ng/ml) of anti-F protein antibodies, determined by ELISA in pooled sera, that binds to the soluble prefusion or postfusion F protein, with time after VLP boosts of RSV-primed animals or a second RSV infection. The values shown are averages with standard deviations from 4 to 5 separate determinations on each of two pools of sera from two different experiments. Values obtained with the two pools of sera from each time point were not statistically significantly different and therefore were combined. (C and D) Levels (ng/ml) of pre-F binding or post-F binding antibodies with time in pools of sera from VLP-primed, VLP-boosted animals. The values shown are averages, with standard deviations, from 4 to 5 separate determinations on each of three pooled sera from three different experiments. Values obtained with each of the three pools of sera were not statistically significantly different and therefore were combined. (E and F) Level (ng/ml) of pre-F (E) or post-F (F) binding antibodies at 4 weeks postboost (the time of maximal NA titers) are shown for clear presentation of statistically significant differences between groups and between the sets of mice. (G and H) Statistically significant differences, at 4 weeks postboost, between pre-F and post-F protein binding antibodies within each set of animals. There were no significant differences between VLP-primed animals at late times postboost. *, P = 0.05; **, P = 0.005; ***, P = 0.0005; ****, P = 0.00005. Only significant differences are indicated.

    Techniques Used: Enzyme-linked Immunosorbent Assay, Infection, Binding Assay, Mouse Assay, Protein Binding

    Splenic B cells secreting anti-F protein antibodies. At weeks 21 and 15 postboost, splenic B cells secreting antibodies that bound soluble prefusion F (A and C) or soluble postfusion F (B and D) were prepared and quantified by ELISpot assay as described in Materials and Methods. Panels show results of duplicate assays for the five animals in each group of RSV-primed, VLP-boosted animals or VLP-primed, VLP-boosted animals. The experiment was repeated twice with two different sets of animals. Panels A and B show results from experiment 1, while panels C and D show results from experiment 2. Statistically significant differences are indicated. *, P = 0.05; **, P = 0.005; ***, P = 0.0005; ****, P = 0.00005.
    Figure Legend Snippet: Splenic B cells secreting anti-F protein antibodies. At weeks 21 and 15 postboost, splenic B cells secreting antibodies that bound soluble prefusion F (A and C) or soluble postfusion F (B and D) were prepared and quantified by ELISpot assay as described in Materials and Methods. Panels show results of duplicate assays for the five animals in each group of RSV-primed, VLP-boosted animals or VLP-primed, VLP-boosted animals. The experiment was repeated twice with two different sets of animals. Panels A and B show results from experiment 1, while panels C and D show results from experiment 2. Statistically significant differences are indicated. *, P = 0.05; **, P = 0.005; ***, P = 0.0005; ****, P = 0.00005.

    Techniques Used: Enzyme-linked Immunospot

    Avidities of anti-F protein antibodies. (A to F) Results with sera from RSV-primed animals. Shown is the stability in increasing concentrations of urea of anti-F antibody-prefusion F protein complexes (A and C) or anti-F antibody-postfusion F protein complexes (D and F) in sera from RSV-primed animals at 0, 4, and 17 weeks after the VLP boost. (A and D) Results using sera from pre-F VLP-boosted animals. (C and F) Results using sera from post-F VLP-boosted animals. (B and E) Results using pre-F VLP sera at 7 M urea in order to illustrate statistically significant differences between groups (*, P = 0.05; **, P = 0.005; ***, P = 0.0005). There were no statistical differences between the values obtained in sera from post-F VLP sera. The results are averages with standard deviations from four separate determinations. RSV/RSV wk 0 indicates RSV/pre-F VLPs at week 0 and RSV/Post-F VLPs at week 0. (G to J) Results with sera from VLP-primed, VLP-boosted animals obtained at 0 and 6 weeks postboost from pre-F VLP (G and I) or post-F VLP (H and J) sera. There were no statistically significant differences between groups of animals at the two time points.
    Figure Legend Snippet: Avidities of anti-F protein antibodies. (A to F) Results with sera from RSV-primed animals. Shown is the stability in increasing concentrations of urea of anti-F antibody-prefusion F protein complexes (A and C) or anti-F antibody-postfusion F protein complexes (D and F) in sera from RSV-primed animals at 0, 4, and 17 weeks after the VLP boost. (A and D) Results using sera from pre-F VLP-boosted animals. (C and F) Results using sera from post-F VLP-boosted animals. (B and E) Results using pre-F VLP sera at 7 M urea in order to illustrate statistically significant differences between groups (*, P = 0.05; **, P = 0.005; ***, P = 0.0005). There were no statistical differences between the values obtained in sera from post-F VLP sera. The results are averages with standard deviations from four separate determinations. RSV/RSV wk 0 indicates RSV/pre-F VLPs at week 0 and RSV/Post-F VLPs at week 0. (G to J) Results with sera from VLP-primed, VLP-boosted animals obtained at 0 and 6 weeks postboost from pre-F VLP (G and I) or post-F VLP (H and J) sera. There were no statistically significant differences between groups of animals at the two time points.

    Techniques Used:

    Experimental protocol. Shown is the timing of RSV, strain A2, infections and VLP immunizations in RSV-primed (infected) (A) and VLP-primed (B) animals. VLP boosts of RSV-primed animals and VLP-primed animals were at 14 weeks postpriming. Upward arrows indicate times of acquisition of serum samples. The first bleed ( t = 0 weeks postboost) was at the time of VLP boosts. At week 21 (RSV primed) or week 15 (VLP primed), animals were infected with RSV, and 4 days later they were sacrificed and terminal bleeds and lungs harvested.
    Figure Legend Snippet: Experimental protocol. Shown is the timing of RSV, strain A2, infections and VLP immunizations in RSV-primed (infected) (A) and VLP-primed (B) animals. VLP boosts of RSV-primed animals and VLP-primed animals were at 14 weeks postpriming. Upward arrows indicate times of acquisition of serum samples. The first bleed ( t = 0 weeks postboost) was at the time of VLP boosts. At week 21 (RSV primed) or week 15 (VLP primed), animals were infected with RSV, and 4 days later they were sacrificed and terminal bleeds and lungs harvested.

    Techniques Used: Infection

    ). RSV infection of animals, sham immunized, served as positive controls. PFU per gram of lung tissue are shown for each mouse in each group of animals. RSV primed, 10 animals/group; VLP primed, 15 animals/group. RSV titers in lungs of each animal were determined.
    Figure Legend Snippet: ). RSV infection of animals, sham immunized, served as positive controls. PFU per gram of lung tissue are shown for each mouse in each group of animals. RSV primed, 10 animals/group; VLP primed, 15 animals/group. RSV titers in lungs of each animal were determined.

    Techniques Used: Infection

    Bone marrow secreting anti-F protein antibodies. At weeks 21 and 15 postboost, bone marrow cells secreting anti-F antibodies that bound soluble prefusion F (A and C) or soluble postfusion F (B and D) were prepared and quantified by ELISpot assay as described in Materials and Methods. Each panel shows results of duplicate assays for the five animals in each group of RSV-primed, VLP-boosted animals or VLP-primed, VLP-boosted animals. The experiment was repeated twice with two different sets of animals. Panels A and B show results from experiment 1, while panels C and D show results from experiment 2. Statistically significant differences are indicated. *, P = 0.05; **, P = 0.005; ***, P = 0.0005; ****, P = 0.00005.
    Figure Legend Snippet: Bone marrow secreting anti-F protein antibodies. At weeks 21 and 15 postboost, bone marrow cells secreting anti-F antibodies that bound soluble prefusion F (A and C) or soluble postfusion F (B and D) were prepared and quantified by ELISpot assay as described in Materials and Methods. Each panel shows results of duplicate assays for the five animals in each group of RSV-primed, VLP-boosted animals or VLP-primed, VLP-boosted animals. The experiment was repeated twice with two different sets of animals. Panels A and B show results from experiment 1, while panels C and D show results from experiment 2. Statistically significant differences are indicated. *, P = 0.05; **, P = 0.005; ***, P = 0.0005; ****, P = 0.00005.

    Techniques Used: Enzyme-linked Immunospot

    25) Product Images from "HSP70L1-mediated intracellular priming of dendritic cell vaccination induces more potent CTL response against cancer"

    Article Title: HSP70L1-mediated intracellular priming of dendritic cell vaccination induces more potent CTL response against cancer

    Journal: Cellular and Molecular Immunology

    doi: 10.1038/cmi.2016.33

    AdCEA 576–669 HSP70L1-DCs induce CEA-specific CD8 +  CTLs. ( a ) The expression of HLA-A11 on LoVo cells with and without β2 m or/plus indicated that nanopeptide (1 μg/ml) was detected using FACS. ( b, c ) Immature DCs, respectively, from HLA-A11 + , HLA-A2.1 +  or HLA-A24 +  healthy donors were pulsed with CEA 576–669 HSP70L1 (CH) or were transfected with indicated Ad for 48 h, then restimulated with autogenous CD3 + T cells for a total of three cycles and then the frequencies of CD8 + T cells specifically recognizing epitopes of HLA-A11-restricted CEA 636–644 , HLA-A2-restricted CAP1 or HLA-A24-restricted CEA 652–660  were detected, respectively, using Pentamer staining and FACS analysis ( b ) or IFN-γ/ELISPOT ( c ), and the cytotoxicity by autogenous CD8 + T cells to LS-174 T, SW480, SW620 or LoVo tumor cells labeled by CFSE was evaluated using cytotoxic assays and FACS analysis ( d ). Representative blots of IFN-γ/ELISPOT ( c , left); HLA-24.2-restricted Her2 263–271 , HLA-A2.1-restricted Her2 435–443  and HLA-A11.1-restricted EBV 416–424  were used as negative control in  c  (right three panels). All results representative of the three independent experiments were shown. Values are % in FACS graphs ( a ,  b ,  d ) mean ±s.d. of three determinants ( c , below). One-way ANOVA ( c ). ** P
    Figure Legend Snippet: AdCEA 576–669 HSP70L1-DCs induce CEA-specific CD8 + CTLs. ( a ) The expression of HLA-A11 on LoVo cells with and without β2 m or/plus indicated that nanopeptide (1 μg/ml) was detected using FACS. ( b, c ) Immature DCs, respectively, from HLA-A11 + , HLA-A2.1 + or HLA-A24 + healthy donors were pulsed with CEA 576–669 HSP70L1 (CH) or were transfected with indicated Ad for 48 h, then restimulated with autogenous CD3 + T cells for a total of three cycles and then the frequencies of CD8 + T cells specifically recognizing epitopes of HLA-A11-restricted CEA 636–644 , HLA-A2-restricted CAP1 or HLA-A24-restricted CEA 652–660 were detected, respectively, using Pentamer staining and FACS analysis ( b ) or IFN-γ/ELISPOT ( c ), and the cytotoxicity by autogenous CD8 + T cells to LS-174 T, SW480, SW620 or LoVo tumor cells labeled by CFSE was evaluated using cytotoxic assays and FACS analysis ( d ). Representative blots of IFN-γ/ELISPOT ( c , left); HLA-24.2-restricted Her2 263–271 , HLA-A2.1-restricted Her2 435–443 and HLA-A11.1-restricted EBV 416–424 were used as negative control in c (right three panels). All results representative of the three independent experiments were shown. Values are % in FACS graphs ( a , b , d ) mean ±s.d. of three determinants ( c , below). One-way ANOVA ( c ). ** P

    Techniques Used: Expressing, FACS, Transfection, Staining, Enzyme-linked Immunospot, Labeling, Negative Control

    The phenotype, allogenetic T-stimulatory activities and inflammatory cytokine secretion by AdCEA 576–669 HSP70L1 DCs. ( a–e ) Immature DCs were pulsed with CEA 576–669 HSP70L1 (CH) or transfected with indicated Ad for 48 h, and then the phenotypes ( a ), allogeneic T-cell-stimulatory activities including the proliferation by CFSE dilution of CD3 + CD4 +  and CD3 + CD8 +  T cells ( b ) and the differentiation of IFN-γ or IL-17-producing CD3 + CD4 + Th cells ( c ) and granzyme B/perforin-producing CD3 + CD8 +  CTL ( d ) and the secretion of IL-6, TNF-α and IL-12/IL-23p40 ( e ) were detected using FACS. Representative results of three independent experiments were shown. Values are % in FACS graphs ( a–d ) or mean±s.e.m of three independent experiments ( e ). * P
    Figure Legend Snippet: The phenotype, allogenetic T-stimulatory activities and inflammatory cytokine secretion by AdCEA 576–669 HSP70L1 DCs. ( a–e ) Immature DCs were pulsed with CEA 576–669 HSP70L1 (CH) or transfected with indicated Ad for 48 h, and then the phenotypes ( a ), allogeneic T-cell-stimulatory activities including the proliferation by CFSE dilution of CD3 + CD4 + and CD3 + CD8 + T cells ( b ) and the differentiation of IFN-γ or IL-17-producing CD3 + CD4 + Th cells ( c ) and granzyme B/perforin-producing CD3 + CD8 + CTL ( d ) and the secretion of IL-6, TNF-α and IL-12/IL-23p40 ( e ) were detected using FACS. Representative results of three independent experiments were shown. Values are % in FACS graphs ( a–d ) or mean±s.e.m of three independent experiments ( e ). * P

    Techniques Used: Transfection, CTL Assay, FACS

    26) Product Images from "In Vivo Efficacy of Glycopeptide-Colistin Combination Therapies in a Galleria mellonella Model of Acinetobacter baumannii Infection ▿"

    Article Title: In Vivo Efficacy of Glycopeptide-Colistin Combination Therapies in a Galleria mellonella Model of Acinetobacter baumannii Infection ▿

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00230-11

    Survival curves for A. baumannii ATCC 19606 (A) and AB210 (B) following treatment with colistin (COL), gentamicin (GENT), teicoplanin (TEIC), or PBS alone. Curves represent a single experiment performed using 16 insects.
    Figure Legend Snippet: Survival curves for A. baumannii ATCC 19606 (A) and AB210 (B) following treatment with colistin (COL), gentamicin (GENT), teicoplanin (TEIC), or PBS alone. Curves represent a single experiment performed using 16 insects.

    Techniques Used:

    Survival of A. baumannii ATCC 19606 (A) and AB210 (B) following treatment with colistin (COL) in combination with teicoplanin (TEIC) or vancomycin (VANC). For ATCC 19606, P
    Figure Legend Snippet: Survival of A. baumannii ATCC 19606 (A) and AB210 (B) following treatment with colistin (COL) in combination with teicoplanin (TEIC) or vancomycin (VANC). For ATCC 19606, P

    Techniques Used:

    27) Product Images from "Polyfunctional Type-1, -2, and -17 CD8+ T Cell Responses to Apoptotic Self-Antigens Correlate with the Chronic Evolution of Hepatitis C Virus Infection"

    Article Title: Polyfunctional Type-1, -2, and -17 CD8+ T Cell Responses to Apoptotic Self-Antigens Correlate with the Chronic Evolution of Hepatitis C Virus Infection

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1002759

    Decay kinetics of pentamer staining for CD8 + T cells. ( A ) Representative decay plot of the natural logarithm of the normalized fluorescence versus time after anti-HLA-A2 mAb addition for fresh PBMCs stained with pentamers that were complexed with the indicated apoptotic epitope. The t 1/2 represents the staining half-times for the pentamers to CD8 + T cells. Filled or empty circles represent PBMCs from patients with acute HCV infection experiencing chronic infection or undergoing infection resolution, respectively. ( B ) The t 1/2 for the pentamers (complexed with apoptotic or viral epitopes) binding CD8 + T cells from patients with acute HCV infection experiencing chronic infection (filled symbols) or undergoing infection resolution (empty symbols). Apoptotic epitopes: circle symbols represent the MYH9 478–485 pentamer specificity, square symbols represent MYH9 741–749 pentamer specificity, and triangle symbols represent VIME 78–87 pentamer specificity. Viral epitopes: circle symbols represent HCV-NS3 1073–1081 pentamer specificity, square symbols represent HCV-NS3 1406–1415 pentamer specificity, and triangle symbols represent HCV-Core 132–140 pentamer specificity.
    Figure Legend Snippet: Decay kinetics of pentamer staining for CD8 + T cells. ( A ) Representative decay plot of the natural logarithm of the normalized fluorescence versus time after anti-HLA-A2 mAb addition for fresh PBMCs stained with pentamers that were complexed with the indicated apoptotic epitope. The t 1/2 represents the staining half-times for the pentamers to CD8 + T cells. Filled or empty circles represent PBMCs from patients with acute HCV infection experiencing chronic infection or undergoing infection resolution, respectively. ( B ) The t 1/2 for the pentamers (complexed with apoptotic or viral epitopes) binding CD8 + T cells from patients with acute HCV infection experiencing chronic infection (filled symbols) or undergoing infection resolution (empty symbols). Apoptotic epitopes: circle symbols represent the MYH9 478–485 pentamer specificity, square symbols represent MYH9 741–749 pentamer specificity, and triangle symbols represent VIME 78–87 pentamer specificity. Viral epitopes: circle symbols represent HCV-NS3 1073–1081 pentamer specificity, square symbols represent HCV-NS3 1406–1415 pentamer specificity, and triangle symbols represent HCV-Core 132–140 pentamer specificity.

    Techniques Used: Staining, Fluorescence, Infection, Binding Assay

    CD8 + T cell multispecificity to apoptotic epitopes in chronic HCV progression. ( A,B ) Mean number of spot-forming cells (SFCs) by fresh CD8 + T effector memory (T EM ) cells (by ELISPOT assay) in response to pools (see Tables S1A–C and S2A–E ) of apoptotic self-epitopes (A) or viral epitopes (B) in HLA-A2 + patients with acute HCV infection experiencing chronic infection (filled circles) or undergoing infection resolution (empty circles), evaluated at different time points of the infection. Each peptide pool contained 5 µg/ml each single peptide. Each circle represents a single patient. NS = not significant. ( C ) Representative correlation (6 th month after clinical onset) between the mean number of SFCs formed by fresh CD8 + T EM cells in response to pools of apoptotic self-epitopes and the viral load (HCV-RNA copies/ml) in patients experiencing chronic infection or undergoing infection resolution. Statistical analysis was performed using non-parametric Spearman's test. ( D ) Representative flow cytometry analyses of PBMCs from a HLA-A2 + patient (Pt.) or a HLA-A2 + healthy donor (H.D.). Cells were then double-stained with a monoclonal antibody (mAb) to CD8 and pentamers expressing the indicated peptides of MYH9, VIME, HCV-NS3, or HCV-Core. Counterplot analyses show the percentage of CD8 + pentamer + cells. The percentage of cells is indicated in each quadrant. ( E,F ) Percentages of CD8 + pentamer + cells from20 HLA-A2 + H.D. and all HLA-A2 + patients studied (filled and empty symbols represent patients experiencing chronic infection or undergoing infection resolution, respectively) evaluated at different time points. In the panel E, circle symbols representMYH9 478–485 pentamer specificity, square symbols represent MYH9 741–749 pentamer specificity, and triangle symbols represent VIME 78–87 pentamer specificity. In the panel F, circle symbols represent HCV-NS3 1073–1081 pentamer specificity, square symbols represent HCV-NS3 1406–1415 pentamer specificity, and triangle symbols represent HCV-Core 132–140 pentamer specificity. NS = not significant. * The percentages of both apoptotic epitope- and viral epitope-specific CD8 + pentamer + cells from H.D. were significantly lower than those from patients (in any time point tested) (p
    Figure Legend Snippet: CD8 + T cell multispecificity to apoptotic epitopes in chronic HCV progression. ( A,B ) Mean number of spot-forming cells (SFCs) by fresh CD8 + T effector memory (T EM ) cells (by ELISPOT assay) in response to pools (see Tables S1A–C and S2A–E ) of apoptotic self-epitopes (A) or viral epitopes (B) in HLA-A2 + patients with acute HCV infection experiencing chronic infection (filled circles) or undergoing infection resolution (empty circles), evaluated at different time points of the infection. Each peptide pool contained 5 µg/ml each single peptide. Each circle represents a single patient. NS = not significant. ( C ) Representative correlation (6 th month after clinical onset) between the mean number of SFCs formed by fresh CD8 + T EM cells in response to pools of apoptotic self-epitopes and the viral load (HCV-RNA copies/ml) in patients experiencing chronic infection or undergoing infection resolution. Statistical analysis was performed using non-parametric Spearman's test. ( D ) Representative flow cytometry analyses of PBMCs from a HLA-A2 + patient (Pt.) or a HLA-A2 + healthy donor (H.D.). Cells were then double-stained with a monoclonal antibody (mAb) to CD8 and pentamers expressing the indicated peptides of MYH9, VIME, HCV-NS3, or HCV-Core. Counterplot analyses show the percentage of CD8 + pentamer + cells. The percentage of cells is indicated in each quadrant. ( E,F ) Percentages of CD8 + pentamer + cells from20 HLA-A2 + H.D. and all HLA-A2 + patients studied (filled and empty symbols represent patients experiencing chronic infection or undergoing infection resolution, respectively) evaluated at different time points. In the panel E, circle symbols representMYH9 478–485 pentamer specificity, square symbols represent MYH9 741–749 pentamer specificity, and triangle symbols represent VIME 78–87 pentamer specificity. In the panel F, circle symbols represent HCV-NS3 1073–1081 pentamer specificity, square symbols represent HCV-NS3 1406–1415 pentamer specificity, and triangle symbols represent HCV-Core 132–140 pentamer specificity. NS = not significant. * The percentages of both apoptotic epitope- and viral epitope-specific CD8 + pentamer + cells from H.D. were significantly lower than those from patients (in any time point tested) (p

    Techniques Used: Enzyme-linked Immunospot, Infection, Flow Cytometry, Cytometry, Staining, Expressing

    28) Product Images from "Binding of TCR Multimers and a TCR-Like Antibody with Distinct Fine-Specificities Is Dependent on the Surface Density of HLA Complexes"

    Article Title: Binding of TCR Multimers and a TCR-Like Antibody with Distinct Fine-Specificities Is Dependent on the Surface Density of HLA Complexes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0051397

    Recognition of pMHC presented on the surface of cells. (A) Untreated (−) and T2 cells treated (+) with IFN-γ were pulsed with 1 or 10 µM of Env 183–191 or 10 µM of Core 18–27 peptide control and stained with 1 µg/mL TCR tetramer. (B) Identically treated T2 cells stained with 1 µg/mL Env183/A2 mAb demonstrates that both treatment with IFN-γ and pulsing with higher peptide concentrations results in a higher surface expression of Env183/A2 complexes. (C) HepG2.2.15 and EBO-PreS1 cells, transfected with the full and PreS1 fragment of the HBV genome, respectively, endogenously process and present Env183/A2 pMHC complexes on their cell surface. The mAb (dashed) shows improved detection compared to the TCR tetramers (solid). Peptide pulsed HepG2.2.15 and EBO-PreS1 further increased surface levels of Env183/A2 (filled histograms). Treatment of cells with IFN-γ demonstrably up-regulated HLA-A2 expression ( Fig. S5B ).
    Figure Legend Snippet: Recognition of pMHC presented on the surface of cells. (A) Untreated (−) and T2 cells treated (+) with IFN-γ were pulsed with 1 or 10 µM of Env 183–191 or 10 µM of Core 18–27 peptide control and stained with 1 µg/mL TCR tetramer. (B) Identically treated T2 cells stained with 1 µg/mL Env183/A2 mAb demonstrates that both treatment with IFN-γ and pulsing with higher peptide concentrations results in a higher surface expression of Env183/A2 complexes. (C) HepG2.2.15 and EBO-PreS1 cells, transfected with the full and PreS1 fragment of the HBV genome, respectively, endogenously process and present Env183/A2 pMHC complexes on their cell surface. The mAb (dashed) shows improved detection compared to the TCR tetramers (solid). Peptide pulsed HepG2.2.15 and EBO-PreS1 further increased surface levels of Env183/A2 (filled histograms). Treatment of cells with IFN-γ demonstrably up-regulated HLA-A2 expression ( Fig. S5B ).

    Techniques Used: Staining, Expressing, Transfection

    29) Product Images from "Bioenergetics of murine lungs infected with respiratory syncytial virus"

    Article Title: Bioenergetics of murine lungs infected with respiratory syncytial virus

    Journal: Virology Journal

    doi: 10.1186/1743-422X-10-22

    Lung respiration in RSV-infected and uninfected BALB/c mice. ( A ) A representative run of O 2 consumption by lung specimen (26 mg) from infected mouse on day 6. The rate of O 2 consumption, k, was set as the negative of the slope of [O 2 ] vs. t (1.4 μM O 2 min -1 ). The value of k after the addition of 10 μM NaCN was 0.3 (78% inhibition), confirming the oxidation occurred mainly in the respiratory chain. Glucose oxidase (catalyzes the reaction of D-glucose + O 2 to D-glucono-δ-lactone + H 2 O 2 ) depleted remaining O 2 in the solution. ( B ) The values of k c (μM O 2 min -1 mg -1 ) for all experiments are plotted as a function of days after inoculation with RSV strain A2 (infected, I) or mock preparation of HEp-2 culture supernatant (uninfected, U). “n” represents number of mice. The data represent at least 10 independent experiments.
    Figure Legend Snippet: Lung respiration in RSV-infected and uninfected BALB/c mice. ( A ) A representative run of O 2 consumption by lung specimen (26 mg) from infected mouse on day 6. The rate of O 2 consumption, k, was set as the negative of the slope of [O 2 ] vs. t (1.4 μM O 2 min -1 ). The value of k after the addition of 10 μM NaCN was 0.3 (78% inhibition), confirming the oxidation occurred mainly in the respiratory chain. Glucose oxidase (catalyzes the reaction of D-glucose + O 2 to D-glucono-δ-lactone + H 2 O 2 ) depleted remaining O 2 in the solution. ( B ) The values of k c (μM O 2 min -1 mg -1 ) for all experiments are plotted as a function of days after inoculation with RSV strain A2 (infected, I) or mock preparation of HEp-2 culture supernatant (uninfected, U). “n” represents number of mice. The data represent at least 10 independent experiments.

    Techniques Used: Infection, Mouse Assay, Inhibition

    30) Product Images from "Mechanism of Action for Respiratory Syncytial Virus Inhibitor RSV604"

    Article Title: Mechanism of Action for Respiratory Syncytial Virus Inhibitor RSV604

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.04119-14

    (A) Effect of RSV604 on intracellular viral replication in HeLa cells. HeLa cells were infected with RSV A2 at an MOI of 0.1 in the presence of DMSO, RSV604 (1× EC 90 ), or AZ-27 (1× EC 90 ). The quantity of RSV RNA in the infected cells was
    Figure Legend Snippet: (A) Effect of RSV604 on intracellular viral replication in HeLa cells. HeLa cells were infected with RSV A2 at an MOI of 0.1 in the presence of DMSO, RSV604 (1× EC 90 ), or AZ-27 (1× EC 90 ). The quantity of RSV RNA in the infected cells was

    Techniques Used: Infection

    Effect of RSV604 on viral RNA synthesis (A), virus release (B), and infectivity (C) in BHK-21 cells. BHK-21 cells were infected with RSV A2 at an MOI of 0.1 in the presence of DMSO (control), RSV604 (1× EC 90 , as in RSV ELISA in HeLa cells) or
    Figure Legend Snippet: Effect of RSV604 on viral RNA synthesis (A), virus release (B), and infectivity (C) in BHK-21 cells. BHK-21 cells were infected with RSV A2 at an MOI of 0.1 in the presence of DMSO (control), RSV604 (1× EC 90 , as in RSV ELISA in HeLa cells) or

    Techniques Used: Infection, Enzyme-linked Immunosorbent Assay

    Effect of RSV604 on virus release and infectivity in HeLa cells. HeLa cells were infected with RSV A2 at an MOI of 0.1 in the presence of DMSO (control), RSV604 (1× EC 90 ), or AZ-27 (1× EC 90 ). (A) The culture supernatants containing the
    Figure Legend Snippet: Effect of RSV604 on virus release and infectivity in HeLa cells. HeLa cells were infected with RSV A2 at an MOI of 0.1 in the presence of DMSO (control), RSV604 (1× EC 90 ), or AZ-27 (1× EC 90 ). (A) The culture supernatants containing the

    Techniques Used: Infection

    (A and B) Potency of RSV604 against RSV A2 infection in HeLa (A) and BHK-21 (B) cells. The data shown are percent inhibition of the RSV signal (means and standard deviations [SD]; n = 3) following a 3-day infection at an MOI of 0.1 as measured by RSV
    Figure Legend Snippet: (A and B) Potency of RSV604 against RSV A2 infection in HeLa (A) and BHK-21 (B) cells. The data shown are percent inhibition of the RSV signal (means and standard deviations [SD]; n = 3) following a 3-day infection at an MOI of 0.1 as measured by RSV

    Techniques Used: Infection, Inhibition

    Effects of RSV604 and AZ-27 on RSV F, N, and M2-1 protein expression and localization. HeLa and BHK-21 cells were infected with RSV A2 at an MOI of 10 in the presence of DMSO (a, b, and c), RSV604 (1× EC 90 , as in RSV ELISA in HeLa cells) (d, e,
    Figure Legend Snippet: Effects of RSV604 and AZ-27 on RSV F, N, and M2-1 protein expression and localization. HeLa and BHK-21 cells were infected with RSV A2 at an MOI of 10 in the presence of DMSO (a, b, and c), RSV604 (1× EC 90 , as in RSV ELISA in HeLa cells) (d, e,

    Techniques Used: Expressing, Infection, Enzyme-linked Immunosorbent Assay

    31) Product Images from "Characterization of a Respiratory Syncytial Virus L Protein Inhibitor"

    Article Title: Characterization of a Respiratory Syncytial Virus L Protein Inhibitor

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.02540-14

    AZ-27 targeting RSV replication. (A) Comparison of the AZ-27 and fusion inhibitor AZD4316 activities in a time-of-addition study. EC 50 s were measured by RSV ELISA in HEp-2 cells following a 3-day infection by RSV A2 at an MOI of 0.02 with compounds added
    Figure Legend Snippet: AZ-27 targeting RSV replication. (A) Comparison of the AZ-27 and fusion inhibitor AZD4316 activities in a time-of-addition study. EC 50 s were measured by RSV ELISA in HEp-2 cells following a 3-day infection by RSV A2 at an MOI of 0.02 with compounds added

    Techniques Used: Enzyme-linked Immunosorbent Assay, Infection

    Susceptibility of AZ-27-resistant virus to RSV inhibitors. RSV A2 virus was cultured in HEp-2 cells in the presence of DMSO (●; control) or AZ-27 (▲, 0.06 μM; □, 0.3 μM) for 4 weeks. The titers of the surviving
    Figure Legend Snippet: Susceptibility of AZ-27-resistant virus to RSV inhibitors. RSV A2 virus was cultured in HEp-2 cells in the presence of DMSO (●; control) or AZ-27 (▲, 0.06 μM; □, 0.3 μM) for 4 weeks. The titers of the surviving

    Techniques Used: Cell Culture

    Discovery of a novel RSV inhibitor. (A) AZ-27 chemical structure. (B) Inhibition of RSV A2 replication by AZ-27. Data shown are percentage of RSV signal (mean ± standard deviation of triplicates) following 3-day infection in HEp-2 cells in the
    Figure Legend Snippet: Discovery of a novel RSV inhibitor. (A) AZ-27 chemical structure. (B) Inhibition of RSV A2 replication by AZ-27. Data shown are percentage of RSV signal (mean ± standard deviation of triplicates) following 3-day infection in HEp-2 cells in the

    Techniques Used: Inhibition, Standard Deviation, Infection

    32) Product Images from "Targeting Hepatitis B Virus-Infected Cells with a T-Cell Receptor-Like Antibody ▿Targeting Hepatitis B Virus-Infected Cells with a T-Cell Receptor-Like Antibody ▿ †"

    Article Title: Targeting Hepatitis B Virus-Infected Cells with a T-Cell Receptor-Like Antibody ▿Targeting Hepatitis B Virus-Infected Cells with a T-Cell Receptor-Like Antibody ▿ †

    Journal: Journal of Virology

    doi: 10.1128/JVI.01990-10

    Characterization of Env183/A2 MAb. (A) Cross-reactivity was evaluated by incubating T2 cells with 1 μM (each) the listed peptides for 1 h, and cells were stained with 0.5 μg of Env183/A2 MAb and analyzed as described above. Specific binding was observed only with the Env183-91 peptide. (B) Env183/A2 MAb is not inhibited by circulating HBV antigens. The Env183/A2 MAb was preincubated with 100 μl of sera from 2 CHB patients, the serum of healthy (AB) subjects, or culture supernatants of HepG2-117 cells. This preincubated MAb-serum mixture was used to stain Env183-91 peptide-loaded (1 μM) T2 cells. (C) Binding characterization of the TCR-like MAb. Titrated concentrations of the Env183/A2 MAb were tested with peptide-pulsed T2 cells. Bars represent the MFI values obtained at the indicated concentrations. Mean values of triplicate measurements are shown. (D) Sensitivity of Env183/A2-specific MAb and CTLs. T2 cells were incubated with the indicated concentrations of the Env183-91 peptide for 1 h and used for the binding assay of antibodies (bars) or CD8 T-cell activation (line). The binding of antibody was detected by flow cytometric analysis as described above. CD8 T-cell activation was calculated as the percentage of CD107-expressing CD8 T cells. Data represent data from one of at least two independent experiments.
    Figure Legend Snippet: Characterization of Env183/A2 MAb. (A) Cross-reactivity was evaluated by incubating T2 cells with 1 μM (each) the listed peptides for 1 h, and cells were stained with 0.5 μg of Env183/A2 MAb and analyzed as described above. Specific binding was observed only with the Env183-91 peptide. (B) Env183/A2 MAb is not inhibited by circulating HBV antigens. The Env183/A2 MAb was preincubated with 100 μl of sera from 2 CHB patients, the serum of healthy (AB) subjects, or culture supernatants of HepG2-117 cells. This preincubated MAb-serum mixture was used to stain Env183-91 peptide-loaded (1 μM) T2 cells. (C) Binding characterization of the TCR-like MAb. Titrated concentrations of the Env183/A2 MAb were tested with peptide-pulsed T2 cells. Bars represent the MFI values obtained at the indicated concentrations. Mean values of triplicate measurements are shown. (D) Sensitivity of Env183/A2-specific MAb and CTLs. T2 cells were incubated with the indicated concentrations of the Env183-91 peptide for 1 h and used for the binding assay of antibodies (bars) or CD8 T-cell activation (line). The binding of antibody was detected by flow cytometric analysis as described above. CD8 T-cell activation was calculated as the percentage of CD107-expressing CD8 T cells. Data represent data from one of at least two independent experiments.

    Techniques Used: Staining, Binding Assay, Incubation, Activation Assay, Flow Cytometry, Expressing

    Schematic representation of the production of Env183/A2 MAb. (A) Synthesis of HLA-A2 complexes, mouse immunization, B-cell selections, and characterization of specific antibody production (detailed in Materials and Methods). The table shows the total number of screened hybridomas and their specificities. (B) Histograms representing the different staining profiles of B-cell hybridoma supernatants. T2 cells were pulsed with 1 μM (each) Env183-91 peptide or influenza A virus matrix peptide at positions 58 to 66 (Flu M1 58-66 ) and then incubated with 50 μl of hybridoma supernatant followed by washing and incubation with anti-mouse IgG-Alexa Fluor 488 for 30 min. Cells were washed and analyzed with flow cytometry. Three different profiles (negative, specific for HLA-A2 molecules, and specific for the Env183/A2 complex) are represented. Data show representative FACS profiles.
    Figure Legend Snippet: Schematic representation of the production of Env183/A2 MAb. (A) Synthesis of HLA-A2 complexes, mouse immunization, B-cell selections, and characterization of specific antibody production (detailed in Materials and Methods). The table shows the total number of screened hybridomas and their specificities. (B) Histograms representing the different staining profiles of B-cell hybridoma supernatants. T2 cells were pulsed with 1 μM (each) Env183-91 peptide or influenza A virus matrix peptide at positions 58 to 66 (Flu M1 58-66 ) and then incubated with 50 μl of hybridoma supernatant followed by washing and incubation with anti-mouse IgG-Alexa Fluor 488 for 30 min. Cells were washed and analyzed with flow cytometry. Three different profiles (negative, specific for HLA-A2 molecules, and specific for the Env183/A2 complex) are represented. Data show representative FACS profiles.

    Techniques Used: Staining, Incubation, Flow Cytometry, Cytometry, FACS

    Fine-mapping of Env183/A2 MAb epitope recognition. (A) Influence of amino acid mutations within the Env183-91 epitope on Env183/A2 MAb and CD8 T-cell recognition. T2 cells were pulsed with 1 μM Env183-91 peptide with or without (wild type [wt]) alanine substitutions at the indicated positions. The inhibition of TCR-like MAb binding (top) and CD8 T-cell activation (bottom) elicited by alanine substitutions on the wild-type Env183-91 peptide are indicated by bars. Percent inhibition is calculated with the following formula: MFI (or CD107 expression) values obtained with alanine-substituted peptides divided by values obtained with wild-type peptides × 100. (B) Env183/A2 MAb is exclusively specific for the Env183-91 peptide of HBV genotypes (gen) A, C, and D. T2 cells were pulsed with 1 μM Env183-91 peptides with the indicated substitutions at position 187. 187R is characteristic of HBV genotype A, C, and D isolates, and K187 is characteristic of HBV genotype B, E, and F isolates. These cells were stained with Env183/A2 MAb for 1 h and analyzed by flow cytometry. Histograms were overlaid. Results are representative of three independent experiments.
    Figure Legend Snippet: Fine-mapping of Env183/A2 MAb epitope recognition. (A) Influence of amino acid mutations within the Env183-91 epitope on Env183/A2 MAb and CD8 T-cell recognition. T2 cells were pulsed with 1 μM Env183-91 peptide with or without (wild type [wt]) alanine substitutions at the indicated positions. The inhibition of TCR-like MAb binding (top) and CD8 T-cell activation (bottom) elicited by alanine substitutions on the wild-type Env183-91 peptide are indicated by bars. Percent inhibition is calculated with the following formula: MFI (or CD107 expression) values obtained with alanine-substituted peptides divided by values obtained with wild-type peptides × 100. (B) Env183/A2 MAb is exclusively specific for the Env183-91 peptide of HBV genotypes (gen) A, C, and D. T2 cells were pulsed with 1 μM Env183-91 peptides with the indicated substitutions at position 187. 187R is characteristic of HBV genotype A, C, and D isolates, and K187 is characteristic of HBV genotype B, E, and F isolates. These cells were stained with Env183/A2 MAb for 1 h and analyzed by flow cytometry. Histograms were overlaid. Results are representative of three independent experiments.

    Techniques Used: Inhibition, Binding Assay, Activation Assay, Expressing, Staining, Flow Cytometry, Cytometry

    33) Product Images from "An AAV Vector-Mediated Gene Delivery Approach Facilitates Reconstitution of Functional Human CD8+ T Cells in Mice"

    Article Title: An AAV Vector-Mediated Gene Delivery Approach Facilitates Reconstitution of Functional Human CD8+ T Cells in Mice

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0088205

    In vitro expression of human cytokines and HLA-A2.1 in MC57G cells. (A) Maps of Zac2.1 plasmids modified to encode human cytokines and HLA-A2.1 (HHD) containing α1 and α2 domains of HLA-A2.1, α3, cytoplasmic and transmembrane domains of murine H-2D b , and hβ2m, are shown. These plasmids were used to construct AAV9 viral particles. (B) MC57G cells were infected in vitro with AAV9 encoding the respective cytokine, and the production of each cytokine was determined using ELISA. (C) MC57G cells were infected in vitro with different doses (1×10 9 , 1×10 10 , or 1×10 11 GC/mL) of AAV9-encoding HLA-A2.1/hβ2m (AAV9-A2). Expression of HLA-A2.1 and hβ2m was evaluated using flow cytometric analyses.
    Figure Legend Snippet: In vitro expression of human cytokines and HLA-A2.1 in MC57G cells. (A) Maps of Zac2.1 plasmids modified to encode human cytokines and HLA-A2.1 (HHD) containing α1 and α2 domains of HLA-A2.1, α3, cytoplasmic and transmembrane domains of murine H-2D b , and hβ2m, are shown. These plasmids were used to construct AAV9 viral particles. (B) MC57G cells were infected in vitro with AAV9 encoding the respective cytokine, and the production of each cytokine was determined using ELISA. (C) MC57G cells were infected in vitro with different doses (1×10 9 , 1×10 10 , or 1×10 11 GC/mL) of AAV9-encoding HLA-A2.1/hβ2m (AAV9-A2). Expression of HLA-A2.1 and hβ2m was evaluated using flow cytometric analyses.

    Techniques Used: In Vitro, Expressing, Modification, Construct, Infection, Enzyme-linked Immunosorbent Assay, Flow Cytometry

    GC number specific for AAV9 and the transgenes present in selected organs of NSG mice at 6 weeks and 20 weeks after AAV9 injection. Lung, liver, spleen, kidney and bone marrow, were collected 6 and 20 weeks after NSG mice were given recombinant AAV9 vectors (5×10 9 GC of AAV9-GM-CSF, AAV9-IL-3, and AAV9-IL-15 by i.v., or 1×10 11 GC of AAV9-A2 by i.v. and intrathoracically), and DNA was isolated from the respective organs. (A) The number of AAV9 vector GC was determined by using a set of primers specific to AAV9 vector. (B) The number of GC specific for the transgenes - HLA-A2, hGM-CSF, hIL-3 and hIL-15 - was determined by using a set of primers to the respective transgene. There was a low number (10–10 3 GC; depending on the transgene) of GC detected in naïve NSG mice, as a non-specific background, and, therefore, this background GC number was subtracted from the GC number in experimental groups.
    Figure Legend Snippet: GC number specific for AAV9 and the transgenes present in selected organs of NSG mice at 6 weeks and 20 weeks after AAV9 injection. Lung, liver, spleen, kidney and bone marrow, were collected 6 and 20 weeks after NSG mice were given recombinant AAV9 vectors (5×10 9 GC of AAV9-GM-CSF, AAV9-IL-3, and AAV9-IL-15 by i.v., or 1×10 11 GC of AAV9-A2 by i.v. and intrathoracically), and DNA was isolated from the respective organs. (A) The number of AAV9 vector GC was determined by using a set of primers specific to AAV9 vector. (B) The number of GC specific for the transgenes - HLA-A2, hGM-CSF, hIL-3 and hIL-15 - was determined by using a set of primers to the respective transgene. There was a low number (10–10 3 GC; depending on the transgene) of GC detected in naïve NSG mice, as a non-specific background, and, therefore, this background GC number was subtracted from the GC number in experimental groups.

    Techniques Used: Mouse Assay, Injection, Recombinant, Isolation, Plasmid Preparation

    In vivo expression of human cytokines and HLA-A2/hβ2-m in NSG mice upon AAV9-mediated gene delivery. (A) NSG mice were inoculated with 5×10 9 GC/mouse of AAV9 encoding each cytokine, and 1, 2, 4, 8, 10, or 16 weeks later, sera were collected from the mice and cytokine production was determined using ELISA. (B) The level of human GM-CSF produced in the sera was determined after inoculation of NSG mice with a high (5×10 9 GC/mouse) or a low (1×10 9 GC/mouse) dose of AAV9-GM-CSF. (C) Luciferase expression 2 weeks after inoculation of NSG mice with 1×10 10 GC of AAV9-GFP-Luc via intrathoracic or i.v. route is shown by injecting D-luciferin intraperitoneally, followed by whole body in vivo imaging analyses. (D) NSG mice were administered intrathoracically with 5×10 10 GC of AAV9-A2 and 4 weeks later, the expression of HLA-A2 and hβ2m by CD326 HIGH cells within the thymus of AAV9-A2-infected NSG mice, A2-Tg NSG mice, and naïve NSG mice was determined using flow cytometric analyses. (E) Immunohistochemical analyses show HLA-A2 (green) and CD326 (red) staining of thymic tissue from AAV9-A2-transduced NSG mice, A2-Tg NSG mice, and naïve NSG mice. Hoechst 33342 (blue) was used to counterstain nuclei.
    Figure Legend Snippet: In vivo expression of human cytokines and HLA-A2/hβ2-m in NSG mice upon AAV9-mediated gene delivery. (A) NSG mice were inoculated with 5×10 9 GC/mouse of AAV9 encoding each cytokine, and 1, 2, 4, 8, 10, or 16 weeks later, sera were collected from the mice and cytokine production was determined using ELISA. (B) The level of human GM-CSF produced in the sera was determined after inoculation of NSG mice with a high (5×10 9 GC/mouse) or a low (1×10 9 GC/mouse) dose of AAV9-GM-CSF. (C) Luciferase expression 2 weeks after inoculation of NSG mice with 1×10 10 GC of AAV9-GFP-Luc via intrathoracic or i.v. route is shown by injecting D-luciferin intraperitoneally, followed by whole body in vivo imaging analyses. (D) NSG mice were administered intrathoracically with 5×10 10 GC of AAV9-A2 and 4 weeks later, the expression of HLA-A2 and hβ2m by CD326 HIGH cells within the thymus of AAV9-A2-infected NSG mice, A2-Tg NSG mice, and naïve NSG mice was determined using flow cytometric analyses. (E) Immunohistochemical analyses show HLA-A2 (green) and CD326 (red) staining of thymic tissue from AAV9-A2-transduced NSG mice, A2-Tg NSG mice, and naïve NSG mice. Hoechst 33342 (blue) was used to counterstain nuclei.

    Techniques Used: In Vivo, Expressing, Mouse Assay, Enzyme-linked Immunosorbent Assay, Produced, Luciferase, In Vivo Imaging, Infection, Flow Cytometry, Immunohistochemistry, Staining

    34) Product Images from "Broad-Spectrum Inhibition of Respiratory Virus Infection by MicroRNA Mimics Targeting p38 MAPK Signaling"

    Article Title: Broad-Spectrum Inhibition of Respiratory Virus Infection by MicroRNA Mimics Targeting p38 MAPK Signaling

    Journal: Molecular Therapy. Nucleic Acids

    doi: 10.1016/j.omtn.2017.03.008

    Reductions in IAV and RSV Titer following Treatments with miRNA Mimics A549 cells were transfected with 25 nM miRNA mimics or controls. A direct-targeting viral siRNA (siIAV or siRSV) served as a positive antiviral control (gray bar), while negative controls were as follows: C. elegans miRNA mimic 1 (C.elegans 1), RISC-free siRNA, Lipofectamine, and virus infection alone (black bars). (A–E) After 48-hr transfection, media was removed and cells were infected with (A) IAV WSN H1N1 at MOI 0.1 for 24 hr, (B) IAV PR8 H1N1 at MOI 0.1 for 12 hr, (C) IAV Udorn H3N2 at MOI 0.1 for 24 hr, (D) RSV-A2 at MOI 0.01 for 72 hr, and (E) RSV-BT2a at MOI 0.1 for 72 hr, when the supernatant was removed and assayed for virus. (F) Uninfected cells were analyzed for cell viability at 48 hr post-transfection (n = 12). The results are displayed to show the most potent antiviral mimics (from left to right along the x axis), with solid lines denoting 0% inhibition, dotted lines denoting the 75% activity cutoff, and blue bars highlighting miRNAs chosen for further analysis. Viral titer results for each virus are shown as the mean ± SEM of n = 6. Significant differences between virus control and miRNA mimics and siRNAs are indicated (*p
    Figure Legend Snippet: Reductions in IAV and RSV Titer following Treatments with miRNA Mimics A549 cells were transfected with 25 nM miRNA mimics or controls. A direct-targeting viral siRNA (siIAV or siRSV) served as a positive antiviral control (gray bar), while negative controls were as follows: C. elegans miRNA mimic 1 (C.elegans 1), RISC-free siRNA, Lipofectamine, and virus infection alone (black bars). (A–E) After 48-hr transfection, media was removed and cells were infected with (A) IAV WSN H1N1 at MOI 0.1 for 24 hr, (B) IAV PR8 H1N1 at MOI 0.1 for 12 hr, (C) IAV Udorn H3N2 at MOI 0.1 for 24 hr, (D) RSV-A2 at MOI 0.01 for 72 hr, and (E) RSV-BT2a at MOI 0.1 for 72 hr, when the supernatant was removed and assayed for virus. (F) Uninfected cells were analyzed for cell viability at 48 hr post-transfection (n = 12). The results are displayed to show the most potent antiviral mimics (from left to right along the x axis), with solid lines denoting 0% inhibition, dotted lines denoting the 75% activity cutoff, and blue bars highlighting miRNAs chosen for further analysis. Viral titer results for each virus are shown as the mean ± SEM of n = 6. Significant differences between virus control and miRNA mimics and siRNAs are indicated (*p

    Techniques Used: Transfection, Infection, Inhibition, Activity Assay

    35) Product Images from "CD4+ T cells support establishment of RSV-specific IgG and IgA antibody secreting cells in the upper and lower murine respiratory tract following RSV infection"

    Article Title: CD4+ T cells support establishment of RSV-specific IgG and IgA antibody secreting cells in the upper and lower murine respiratory tract following RSV infection

    Journal: Vaccine

    doi: 10.1016/j.vaccine.2017.03.073

    Serum antibodies in RSV-infected mice Nude mice were infected with RSV and 14 days later received no cells, splenocytes from RSV-primed BALB/c mice enriched for CD4 + T cells (depleted of CD8 + and MHC class II + cells including B cells, monocytes, macrophages, and dendritic cells [‘CD4s’]), or the same splenocytes additionally depleted of CD4 + T cells (‘Depleted CD4s’). Antibody responses against RSV or a purified RSV fusion protein (F) were analyzed by ELISAs 5 weeks after RSV infections.
    Figure Legend Snippet: Serum antibodies in RSV-infected mice Nude mice were infected with RSV and 14 days later received no cells, splenocytes from RSV-primed BALB/c mice enriched for CD4 + T cells (depleted of CD8 + and MHC class II + cells including B cells, monocytes, macrophages, and dendritic cells [‘CD4s’]), or the same splenocytes additionally depleted of CD4 + T cells (‘Depleted CD4s’). Antibody responses against RSV or a purified RSV fusion protein (F) were analyzed by ELISAs 5 weeks after RSV infections.

    Techniques Used: Infection, Mouse Assay, Purification

    36) Product Images from "Novel Rhamnosyltransferase Involved in Biosynthesis of Serovar 4-Specific Glycopeptidolipid from Mycobacterium avium Complex ▿"

    Article Title: Novel Rhamnosyltransferase Involved in Biosynthesis of Serovar 4-Specific Glycopeptidolipid from Mycobacterium avium Complex ▿

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00554-10

    Organization of the 6.8-kb genomic segment isolated from MAC serovar 4 strain (ATCC 35767). Filled triangles indicate the primers used for PCR amplification.
    Figure Legend Snippet: Organization of the 6.8-kb genomic segment isolated from MAC serovar 4 strain (ATCC 35767). Filled triangles indicate the primers used for PCR amplification.

    Techniques Used: Isolation, Polymerase Chain Reaction, Amplification

    37) Product Images from "Both CD4 and CD8 T Cells Mediate Equally Effective In Vivo Tumor Treatment When Engineered with a Highly Avid TCR Targeting Tyrosinase"

    Article Title: Both CD4 and CD8 T Cells Mediate Equally Effective In Vivo Tumor Treatment When Engineered with a Highly Avid TCR Targeting Tyrosinase

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1000189

    Transduction of mouse splenocytes with the antityrosinase TCR confers peptide reactivity and tumor recognition. A , Isotype, Vβ12, and CD8 staining of splenocytes following 48 h stimulation in OKT3 and CD28 and a single transduction with retrovirus encoding the antityrosinase TCR. B , IFN-γ release following coculture with T2 cells pulsed with decreasing concentrations of peptide ( left panel ) and tumor targets ( right panel ) B16/A2/K b (HLA-A2 + , tyrosinase + ), B16 wild-type (HLA-A2 − , tyrosinase + ), 526mel, 624mel, and 888mel.
    Figure Legend Snippet: Transduction of mouse splenocytes with the antityrosinase TCR confers peptide reactivity and tumor recognition. A , Isotype, Vβ12, and CD8 staining of splenocytes following 48 h stimulation in OKT3 and CD28 and a single transduction with retrovirus encoding the antityrosinase TCR. B , IFN-γ release following coculture with T2 cells pulsed with decreasing concentrations of peptide ( left panel ) and tumor targets ( right panel ) B16/A2/K b (HLA-A2 + , tyrosinase + ), B16 wild-type (HLA-A2 − , tyrosinase + ), 526mel, 624mel, and 888mel.

    Techniques Used: Transduction, Staining

    38) Product Images from "High Therapeutic Efficacy of a New Survivin LSP-Cancer Vaccine Containing CD4+ and CD8+ T-Cell Epitopes"

    Article Title: High Therapeutic Efficacy of a New Survivin LSP-Cancer Vaccine Containing CD4+ and CD8+ T-Cell Epitopes

    Journal: Frontiers in Oncology

    doi: 10.3389/fonc.2018.00517

    High therapeutic efficacy of SVX vaccine against various established tumor models . (A) T-cell immunogenicity of SVX vaccine and adjuvant selection. Tumor-free BALB/c (H2 d ) mice were subcutaneously (s.c) vaccinated as followed: priming with the three survivin LSPs (SVX) and adjuvants and boost 2 weeks later with SVX without adjuvant. One week after the boost, the induction of survivin-specific T-cell responses was analyzed by IFN-γ ELISpot assay on total splenocytes (2 × 10 5 cells). Left and middle graphs showed overnight restimulation with the pool of SVX peptides or individual peptides ( A , left graph) or tumor cell lines expressing the human survivin (hCT26 and hA20) ( A , middle graph). Comparison of SVX specific T-cell responses induced by various adjuvants was performed by restimulation with the pool of SVX peptides ( A , right graph). Results are the mean ± SEM of 5 mice per group and are representative of two to three independent experiments. *** P
    Figure Legend Snippet: High therapeutic efficacy of SVX vaccine against various established tumor models . (A) T-cell immunogenicity of SVX vaccine and adjuvant selection. Tumor-free BALB/c (H2 d ) mice were subcutaneously (s.c) vaccinated as followed: priming with the three survivin LSPs (SVX) and adjuvants and boost 2 weeks later with SVX without adjuvant. One week after the boost, the induction of survivin-specific T-cell responses was analyzed by IFN-γ ELISpot assay on total splenocytes (2 × 10 5 cells). Left and middle graphs showed overnight restimulation with the pool of SVX peptides or individual peptides ( A , left graph) or tumor cell lines expressing the human survivin (hCT26 and hA20) ( A , middle graph). Comparison of SVX specific T-cell responses induced by various adjuvants was performed by restimulation with the pool of SVX peptides ( A , right graph). Results are the mean ± SEM of 5 mice per group and are representative of two to three independent experiments. *** P

    Techniques Used: Selection, Mouse Assay, Enzyme-linked Immunospot, Expressing

    Generation of long term anti-tumor memory T cells following SVX vaccination. (A–C) Studies on tumor regression. (A) Histograms represent the percentage ± SEM of complete tumor regression of a pool of five different experiments (representing 40 mice) for both hCT26 and hA20 tumor models. (B) A group of vaccinated mice engrafted with hA20 cells ( n = 5), which completely eliminated tumor cells were re-challenged with hA20 tumor cells. As positive control naive BALB/c mice ( n = 5) were engrafted with hA20 tumor cells (Naive mice). Tumor growth (B) and Survival (C) were monitored. The experiment has been performed twice. ** P
    Figure Legend Snippet: Generation of long term anti-tumor memory T cells following SVX vaccination. (A–C) Studies on tumor regression. (A) Histograms represent the percentage ± SEM of complete tumor regression of a pool of five different experiments (representing 40 mice) for both hCT26 and hA20 tumor models. (B) A group of vaccinated mice engrafted with hA20 cells ( n = 5), which completely eliminated tumor cells were re-challenged with hA20 tumor cells. As positive control naive BALB/c mice ( n = 5) were engrafted with hA20 tumor cells (Naive mice). Tumor growth (B) and Survival (C) were monitored. The experiment has been performed twice. ** P

    Techniques Used: Mouse Assay, Positive Control

    SVX therapeutic efficacy against tumor cells in CD8-depleted mice. BALB/c mice (8 mice per group) were engrafted s.c with hCT26 (A,C) or hA20 tumor cells (B,D) . When tumors reached 10 mm 2 , mice were s.c injected with PBS, or were immunized with SVX + CpG/IFA and received a boost 1 week later without adjuvant (SVX). (A,B) Groups of vaccinated mice engrafted with hCT26 (A) or hA20 tumor cells (B) were depleted of CD8 + T cells, using anti-CD8 mAbs (100 μg) injected intra-peritoneally (i.p) once a week, starting 1 day before SVX immunization (SVX + αCD8). Data are presented as mean tumor size (mm 2 ) ± SEM from cohorts of 8 mice with * P
    Figure Legend Snippet: SVX therapeutic efficacy against tumor cells in CD8-depleted mice. BALB/c mice (8 mice per group) were engrafted s.c with hCT26 (A,C) or hA20 tumor cells (B,D) . When tumors reached 10 mm 2 , mice were s.c injected with PBS, or were immunized with SVX + CpG/IFA and received a boost 1 week later without adjuvant (SVX). (A,B) Groups of vaccinated mice engrafted with hCT26 (A) or hA20 tumor cells (B) were depleted of CD8 + T cells, using anti-CD8 mAbs (100 μg) injected intra-peritoneally (i.p) once a week, starting 1 day before SVX immunization (SVX + αCD8). Data are presented as mean tumor size (mm 2 ) ± SEM from cohorts of 8 mice with * P

    Techniques Used: Mouse Assay, Injection, Immunofluorescence

    39) Product Images from "Recognition of NY-ESO-1+ tumor cells by engineered lymphocytes is enhanced by improved vector design and epigenetic modulation of tumor antigen expression"

    Article Title: Recognition of NY-ESO-1+ tumor cells by engineered lymphocytes is enhanced by improved vector design and epigenetic modulation of tumor antigen expression

    Journal: Cancer immunology, immunotherapy : CII

    doi: 10.1007/s00262-008-0562-x

    Cytokine production of TCR-transduced PBLs following co-culture with NY-ESO-1+, HLA-A2+ and − cell lines. PBL were transduced with supernatant from the CysfrnSGSGP2A retroviral construct and were then co-cultured with the H1299 non-small cell lung carcinoma line (HLA-A2-transfected) (NY-ESO-1+, HLA-A2+), the (BE-3 esophageal carcinoma cell line (NY-ESO-1−, HLA-A2+), the H2373 pleural mesothelioma cell line (NY-ESO-1−, HLA-A2+),), the human pancreatic cancer cell line Panc-1 (NY-ESO-1−, HLA-A2+), the human osteosarcoma cell line LNZTA3WT4 (NY-ESO-1−, HLA-A2+), and the human ovarian cancer cell line OVCAR-3 (NY-ESO-1−, HLA-A2+) after exposure to normal media (NM), 5-aza-2′-deoxycytidine (DAC), depsipeptide (DP), or sequential 5-aza-2′-deoxycytidine and depsipeptide (DAC-DP). Antigen-specific IFN- γ secretion by retroviral vector-transduced PBLs was determined by ELISA. Data are the mean values (picograms per milliliter) of duplicate samples, with standard deviation ( error bars )
    Figure Legend Snippet: Cytokine production of TCR-transduced PBLs following co-culture with NY-ESO-1+, HLA-A2+ and − cell lines. PBL were transduced with supernatant from the CysfrnSGSGP2A retroviral construct and were then co-cultured with the H1299 non-small cell lung carcinoma line (HLA-A2-transfected) (NY-ESO-1+, HLA-A2+), the (BE-3 esophageal carcinoma cell line (NY-ESO-1−, HLA-A2+), the H2373 pleural mesothelioma cell line (NY-ESO-1−, HLA-A2+),), the human pancreatic cancer cell line Panc-1 (NY-ESO-1−, HLA-A2+), the human osteosarcoma cell line LNZTA3WT4 (NY-ESO-1−, HLA-A2+), and the human ovarian cancer cell line OVCAR-3 (NY-ESO-1−, HLA-A2+) after exposure to normal media (NM), 5-aza-2′-deoxycytidine (DAC), depsipeptide (DP), or sequential 5-aza-2′-deoxycytidine and depsipeptide (DAC-DP). Antigen-specific IFN- γ secretion by retroviral vector-transduced PBLs was determined by ELISA. Data are the mean values (picograms per milliliter) of duplicate samples, with standard deviation ( error bars )

    Techniques Used: Co-Culture Assay, Transduction, Construct, Cell Culture, Transfection, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Standard Deviation

    Design of NY-ESO-1 TCR retroviral vectors. Schematic representation of six retroviral vectors used to transfer and express the TCR genes targeting the HLA-A2-restricted immundominant epitope of NY-ESO-1. Vector MSGV1-1G4-AIB is a bicistronic vector design where the LTR promoter drives the expression of both TCR α and β genes linked by an IRES. In vector MSGV1-1G4-APB (APB), the expression of the α chain is mediated by the vector LTR gene promoter, whereas β chain expression is driven by an internal PGK promoter. In each of the vectors incorporating 2A linker peptides, the expression of both the α and β chains is mediated by the vector LTR gene promoter through an open reading frame, and ‘cleavage’ of the α - and β chains is accomplished at the protein level, where incomplete peptide bond formation between two components in the linker peptide design results in two separate proteins. MSGV1-1G4-AGSGP2AB (GSGP2A) incorporates coding sequences generated from the porcine teschovirus-1 and includes a GSG (glycine-serine-glycine) ‘spacer’ sequence. MSGV1-1G4-AGSGT2AB (GSGT2A) incorporates coding sequences generated from the Thosea asigna virus and includes a GSG ‘spacer’ sequence. MSGV1-1G4-AT2AB (T2A) is essentially identical to MSGV1-1G4-AGSGT2AB with the exception that it lacks a GSG ‘spacer’ sequence. The MSGV1-Cys1G4-AfrnSGSGP2AB (Cys-frnSGSGP2A) vector incorporates α and β chains of the 1G4 NY-ESO-1 TCR in which we replaced a Thr 48 on the α chain and a Ser 57 on the β chain with cysteines to facilitate the creation of an additional disulfide bond between the TCR constant regions in an effort to enhance TCR stability
    Figure Legend Snippet: Design of NY-ESO-1 TCR retroviral vectors. Schematic representation of six retroviral vectors used to transfer and express the TCR genes targeting the HLA-A2-restricted immundominant epitope of NY-ESO-1. Vector MSGV1-1G4-AIB is a bicistronic vector design where the LTR promoter drives the expression of both TCR α and β genes linked by an IRES. In vector MSGV1-1G4-APB (APB), the expression of the α chain is mediated by the vector LTR gene promoter, whereas β chain expression is driven by an internal PGK promoter. In each of the vectors incorporating 2A linker peptides, the expression of both the α and β chains is mediated by the vector LTR gene promoter through an open reading frame, and ‘cleavage’ of the α - and β chains is accomplished at the protein level, where incomplete peptide bond formation between two components in the linker peptide design results in two separate proteins. MSGV1-1G4-AGSGP2AB (GSGP2A) incorporates coding sequences generated from the porcine teschovirus-1 and includes a GSG (glycine-serine-glycine) ‘spacer’ sequence. MSGV1-1G4-AGSGT2AB (GSGT2A) incorporates coding sequences generated from the Thosea asigna virus and includes a GSG ‘spacer’ sequence. MSGV1-1G4-AT2AB (T2A) is essentially identical to MSGV1-1G4-AGSGT2AB with the exception that it lacks a GSG ‘spacer’ sequence. The MSGV1-Cys1G4-AfrnSGSGP2AB (Cys-frnSGSGP2A) vector incorporates α and β chains of the 1G4 NY-ESO-1 TCR in which we replaced a Thr 48 on the α chain and a Ser 57 on the β chain with cysteines to facilitate the creation of an additional disulfide bond between the TCR constant regions in an effort to enhance TCR stability

    Techniques Used: Plasmid Preparation, Expressing, Generated, Sequencing

    Cytokine production of TCR-transduced PBLs following co-culture with cells expressing the cognate antigen. a PBL transduced with supernatant from packaging cell clones APB 38, GSGP2A 3, GSGT2A 36, and T2A 24 were co-cultured with T2 cells pulsed with NY-ESO-1 peptide (p157–165v) or influenza-specific peptide. Antigen-specific IFN- γ secretion by retroviral vector-transduced PBLs was determined by ELISA. Data are the mean values (picograms per milliliter) of duplicate samples, with standard deviation ( error bars ). b PBL were transduced with supernatant from retroviral constructs for GFP (control virus) or from APB, GSGP2A, GSGT2A, and T2A and were then co-cultured with HLA-A2+, NYESO-1+ melanoma lines (624.38mel, 1300mel, A375mel) or HLA-A2+, NY-ESO-1- melanoma line (SK23). Transduced PBL were also incubated with T2 cells alone, or T2 cells pulsed with NY-ESO-1 peptide (p157–165v) or gp100 peptide. Antigen-specific IFN- γ secretion by retroviral vector-transduced PBLs was determined by ELISA. Data are the mean values (picograms per milliliter) of samples, with standard deviation ( error bars )
    Figure Legend Snippet: Cytokine production of TCR-transduced PBLs following co-culture with cells expressing the cognate antigen. a PBL transduced with supernatant from packaging cell clones APB 38, GSGP2A 3, GSGT2A 36, and T2A 24 were co-cultured with T2 cells pulsed with NY-ESO-1 peptide (p157–165v) or influenza-specific peptide. Antigen-specific IFN- γ secretion by retroviral vector-transduced PBLs was determined by ELISA. Data are the mean values (picograms per milliliter) of duplicate samples, with standard deviation ( error bars ). b PBL were transduced with supernatant from retroviral constructs for GFP (control virus) or from APB, GSGP2A, GSGT2A, and T2A and were then co-cultured with HLA-A2+, NYESO-1+ melanoma lines (624.38mel, 1300mel, A375mel) or HLA-A2+, NY-ESO-1- melanoma line (SK23). Transduced PBL were also incubated with T2 cells alone, or T2 cells pulsed with NY-ESO-1 peptide (p157–165v) or gp100 peptide. Antigen-specific IFN- γ secretion by retroviral vector-transduced PBLs was determined by ELISA. Data are the mean values (picograms per milliliter) of samples, with standard deviation ( error bars )

    Techniques Used: Co-Culture Assay, Expressing, Transduction, Clone Assay, Cell Culture, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Standard Deviation, Construct, Incubation

    T cell receptor expression and effector function of PBL transduced with modified TCR vector. a A retroviral construct incorporating cysteine-modified α and β chains was constructed as previously described. OKT3-stimulated PBL were transduced with viral supernatant from the GSGP2A and CysfrnSGSGP2A constructs. Five days post transduction, PBL were stained for V β 13.1 and NY-ESO-1 tetramer and expression was analyzed via flow cytometry. The percentage of positive cells for V β 13.1 and NY-ESO-1 are indicated on the histogram. b PBL were transduced with supernatant from retroviral constructs for GFP (control virus) or from GSGP2A and CysfrnSGSGP2A and were then co-cultured with HLA-A2+, NYESO-1+ melanoma lines (624.38mel, 1300mel, A375mel) or HLA-A2+, NY-ESO-1− melanoma line (SK23). Antigen-specific IFN- γ secretion by retroviral vector-transduced PBLs was determined by ELISA. Data are the mean values (picograms per milliliter) of samples, with standard deviation ( error bars )
    Figure Legend Snippet: T cell receptor expression and effector function of PBL transduced with modified TCR vector. a A retroviral construct incorporating cysteine-modified α and β chains was constructed as previously described. OKT3-stimulated PBL were transduced with viral supernatant from the GSGP2A and CysfrnSGSGP2A constructs. Five days post transduction, PBL were stained for V β 13.1 and NY-ESO-1 tetramer and expression was analyzed via flow cytometry. The percentage of positive cells for V β 13.1 and NY-ESO-1 are indicated on the histogram. b PBL were transduced with supernatant from retroviral constructs for GFP (control virus) or from GSGP2A and CysfrnSGSGP2A and were then co-cultured with HLA-A2+, NYESO-1+ melanoma lines (624.38mel, 1300mel, A375mel) or HLA-A2+, NY-ESO-1− melanoma line (SK23). Antigen-specific IFN- γ secretion by retroviral vector-transduced PBLs was determined by ELISA. Data are the mean values (picograms per milliliter) of samples, with standard deviation ( error bars )

    Techniques Used: Expressing, Transduction, Modification, Plasmid Preparation, Construct, Staining, Flow Cytometry, Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay, Standard Deviation

    40) Product Images from "Identification of MAGE-C1 (CT-7) epitopes for T-cell therapy of multiple myeloma"

    Article Title: Identification of MAGE-C1 (CT-7) epitopes for T-cell therapy of multiple myeloma

    Journal: Cancer immunology, immunotherapy : CII

    doi: 10.1007/s00262-011-1009-3

    Expression of CT-7 by myeloma cells. Myeloma cell lines (H929, U266, and L363) and bone marrow samples from 11 patients with stage II–III multiple myeloma (MM) were stained for intracellular CT-7 expression ( gray histograms ) or with an isotype
    Figure Legend Snippet: Expression of CT-7 by myeloma cells. Myeloma cell lines (H929, U266, and L363) and bone marrow samples from 11 patients with stage II–III multiple myeloma (MM) were stained for intracellular CT-7 expression ( gray histograms ) or with an isotype

    Techniques Used: Expressing, Staining

    Related Articles

    Isolation:

    Article Title: Characterization of the Antibody Response to the Receptor Binding Domain of Botulinum Neurotoxin Serotypes A and E
    Article Snippet: .. Sera isolated from mice and rabbits immunized with recombinant HCR/A1 (rHCR/A1) from the classical type A-Hall strain (ATCC 3502) (BoNT/A1) and rHCR/E from BoNT serotype E Beluga (BoNT/EB ) neutralized the homologous serotype of BoNT but displayed differences in cross-recognition and cross-protection. .. Enzyme-linked immunosorbent assay and Western blotting showed that α-rHCR/A1 recognized epitopes within the C terminus of the HCR/A and HCR/E, while α-rHCR/E recognized epitopes within the N terminus or interface between the N and C termini of the HCR proteins. α-rHCR/EB sera possessed detectable neutralizing capacity for BoNT/A1, while α-rHCR/A1 did not neutralize BoNT/E. rHCR/A was an effective immunogen against BoNT/A1 and the Kyoto F infant strain (BoNT/A2), but not BoNT serotype E Alaska (BoNT/EA ), while rHCR/EB neutralized BoNT/EA , and under hyperimmunization conditions protected against BoNT/A1 and BoNT/A2.

    Infection:

    Article Title: Combination Therapy Using Monoclonal Antibodies against Respiratory Syncytial Virus (RSV) G Glycoprotein Protects from RSV Disease in BALB/c Mice
    Article Snippet: .. Virus Infection and Tissue Collection The A2 strain of RSV was used in all experiments and propagated in Vero cells (ATCC CCL 881) as previously described . .. Mice were anesthetized by intraperitoneally (i.p.) administration of Avertin (2% 2,2,2-tribromoethanol, 2% tert-amyl-alcohol, 180–250 mg/kg), and intranasally (i.n.) challenged with 106 plaques forming units (PFU) of RSV in serum free DMEM in a 50 µL volume.

    Mouse Assay:

    Article Title: Characterization of the Antibody Response to the Receptor Binding Domain of Botulinum Neurotoxin Serotypes A and E
    Article Snippet: .. Sera isolated from mice and rabbits immunized with recombinant HCR/A1 (rHCR/A1) from the classical type A-Hall strain (ATCC 3502) (BoNT/A1) and rHCR/E from BoNT serotype E Beluga (BoNT/EB ) neutralized the homologous serotype of BoNT but displayed differences in cross-recognition and cross-protection. .. Enzyme-linked immunosorbent assay and Western blotting showed that α-rHCR/A1 recognized epitopes within the C terminus of the HCR/A and HCR/E, while α-rHCR/E recognized epitopes within the N terminus or interface between the N and C termini of the HCR proteins. α-rHCR/EB sera possessed detectable neutralizing capacity for BoNT/A1, while α-rHCR/A1 did not neutralize BoNT/E. rHCR/A was an effective immunogen against BoNT/A1 and the Kyoto F infant strain (BoNT/A2), but not BoNT serotype E Alaska (BoNT/EA ), while rHCR/EB neutralized BoNT/EA , and under hyperimmunization conditions protected against BoNT/A1 and BoNT/A2.

    Generated:

    Article Title: Diagnostic utility of LunX mRNA in peripheral blood and pleural fluid in patients with primary non-small cell lung cancer
    Article Snippet: .. Preparation of standard curves LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin cDNA were generated from A549, SK-BR-3, SK-BR-3, K562, A549 and A549 cells (American Type Culture Collection, Rockville, MD), respectively, using the specific primers in Table . .. The PCR reaction consisted of an initial denaturation step at 94°C for 3 min, followed by 35 cycles of denaturation at 94°C, annealing at 60°C for CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin or at 56°C for LunX , and extension at 72°C.

    Sequencing:

    Article Title: Characterization of a potential ABC-type bacteriocin exporter protein from Treponema denticola
    Article Snippet: .. A search of the flanking regions of tepA1 , A2 , and A3 in the genome sequence of T. denticola ATCC 35405 revealed that proteins coded by three open reading frames (TDE_0416, TDE_0422, and TDE_0423) upstream of tepA1 have double-glycine bacteriocin-type signal domains. .. However, no double glycine-containing protein-coding sequence exists near tepA2 and A3.

    Recombinant:

    Article Title: Characterization of the Antibody Response to the Receptor Binding Domain of Botulinum Neurotoxin Serotypes A and E
    Article Snippet: .. Sera isolated from mice and rabbits immunized with recombinant HCR/A1 (rHCR/A1) from the classical type A-Hall strain (ATCC 3502) (BoNT/A1) and rHCR/E from BoNT serotype E Beluga (BoNT/EB ) neutralized the homologous serotype of BoNT but displayed differences in cross-recognition and cross-protection. .. Enzyme-linked immunosorbent assay and Western blotting showed that α-rHCR/A1 recognized epitopes within the C terminus of the HCR/A and HCR/E, while α-rHCR/E recognized epitopes within the N terminus or interface between the N and C termini of the HCR proteins. α-rHCR/EB sera possessed detectable neutralizing capacity for BoNT/A1, while α-rHCR/A1 did not neutralize BoNT/E. rHCR/A was an effective immunogen against BoNT/A1 and the Kyoto F infant strain (BoNT/A2), but not BoNT serotype E Alaska (BoNT/EA ), while rHCR/EB neutralized BoNT/EA , and under hyperimmunization conditions protected against BoNT/A1 and BoNT/A2.

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    ATCC hnrnp a2 b1
    LunX mRNA is the most specific gene marker for lung cancer cells in peripheral blood . LunX, CK19, CEA, VEGF-C, <t>hnRNP</t> <t>A2/B1</t> and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin in all samples. For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as 50%, the median cannot be shown. " width="250" height="auto" />
    Hnrnp A2 B1, supplied by ATCC, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hnrnp a2 b1/product/ATCC
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    91
    ATCC prototype a2 strain
    Neutralizing antibody response following RSV-G protein, FI-RSV and live RSV experimental infection. ( A ) Schematic representation of cotton rat immunization and challenge schedule. Inbred female  Sigmodon hispidus  cotton rats between 6 and 8 weeks of age were immunized i.m. with PBS (Gps A-B), 5 μg of unadjuvanted or Emulsigen-adjuvanted REG ( R ecombinant  E . coli  produced  G ) (Gps C-D), or with FI-RSV (Gp E) on days 0 and 28 in groups A thru E, or were infected intranasally (i.n.) with 0.1 ml of RSV/A2 at 10 5  pfu per rat (Gp F). Blood was collected by eye-bleed on days 0, 28 and 49. On day 49, animals were either mock challenge intranasally with 0.1 ml of PBS (Gp A), or with 0.1 ml of RSV-A2 virus at 10 5  pfu per animal (10 animals per group) (Gps B-F). Cotton rats were sacrificed on days 2 or 5 post-challenge wherein lungs and nose tissues were collected. ( B ) Sera from individual cotton rats collected at pre-vaccination (day 0) and 3 weeks post second immunization (day 49) were tested for neutralization in a plaque reduction neutralization test (PRNT) against the homologous RSV-A2 strain and heterologous RSV-B1 strain. Neutralizing antibody titers represent 50% inhibition of plaque numbers. Statistical significance was tested with one-way ANOVA and Bonferroni multiple comparisons tests. ***p 
    Prototype A2 Strain, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    ATCC authentic gentamicin a2
    GtmE Acts as an UDP-Xylose Glycosyltransferase to Generate <t>Gentamicin</t> A2 (4a).
    Authentic Gentamicin A2, supplied by ATCC, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/authentic gentamicin a2/product/ATCC
    Average 85 stars, based on 1 article reviews
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    authentic gentamicin a2 - by Bioz Stars, 2020-09
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    85
    ATCC strains abh12o a2
    PCR fingerprinting profile of plasmids pMMA2 and pMCU3, determined by using a set of oligonucleotides. Lane 1, PCR fingerprint of plasmids isolated from strain <t>AbH12O-A2;</t> lane 2, PCR fingerprint of plasmids isolated from strain ATCC 17978 transformed
    Strains Abh12o A2, supplied by ATCC, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/strains abh12o a2/product/ATCC
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    LunX mRNA is the most specific gene marker for lung cancer cells in peripheral blood . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin in all samples. For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as 50%, the median cannot be shown. " width="100%" height="100%">

    Journal: BMC Cancer

    Article Title: Diagnostic utility of LunX mRNA in peripheral blood and pleural fluid in patients with primary non-small cell lung cancer

    doi: 10.1186/1471-2407-8-156

    Figure Lengend Snippet: LunX mRNA is the most specific gene marker for lung cancer cells in peripheral blood . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin in all samples. For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as " --- " in each group. Where the frequency of negative cases is > 50%, the median cannot be shown.

    Article Snippet: Preparation of standard curves LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin cDNA were generated from A549, SK-BR-3, SK-BR-3, K562, A549 and A549 cells (American Type Culture Collection, Rockville, MD), respectively, using the specific primers in Table .

    Techniques: Marker, Quantitative RT-PCR

    Expression of LunX mRNA in peripheral blood decreases shortly following the treatment of NSCLC . Peripheral blood samples from 12 NSCLC patients were collected 1 day before and 7 days after treatment as shown in Table 2. LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods.

    Journal: BMC Cancer

    Article Title: Diagnostic utility of LunX mRNA in peripheral blood and pleural fluid in patients with primary non-small cell lung cancer

    doi: 10.1186/1471-2407-8-156

    Figure Lengend Snippet: Expression of LunX mRNA in peripheral blood decreases shortly following the treatment of NSCLC . Peripheral blood samples from 12 NSCLC patients were collected 1 day before and 7 days after treatment as shown in Table 2. LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. "Tb" represents 1 day before treatment and "Ta" represents 7 days after treatment. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The Wilcoxon Signed Ranks Test was used to analyze the gene expression levels before and after clinical treatment. P

    Article Snippet: Preparation of standard curves LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin cDNA were generated from A549, SK-BR-3, SK-BR-3, K562, A549 and A549 cells (American Type Culture Collection, Rockville, MD), respectively, using the specific primers in Table .

    Techniques: Expressing, Quantitative RT-PCR, Marker

    Expression level of LunX mRNA in peripheral blood is correlated with the pathologic stage of NSCLC . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as 50%, the median cannot be shown. The K Independent Samples Test (Media Test) was used to analyze gene expression levels among NSCLC patients at different pathologic stages. P " width="100%" height="100%">

    Journal: BMC Cancer

    Article Title: Diagnostic utility of LunX mRNA in peripheral blood and pleural fluid in patients with primary non-small cell lung cancer

    doi: 10.1186/1471-2407-8-156

    Figure Lengend Snippet: Expression level of LunX mRNA in peripheral blood is correlated with the pathologic stage of NSCLC . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each peripheral blood sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as " --- " in each group. Where the frequency of negative cases is > 50%, the median cannot be shown. The K Independent Samples Test (Media Test) was used to analyze gene expression levels among NSCLC patients at different pathologic stages. P

    Article Snippet: Preparation of standard curves LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin cDNA were generated from A549, SK-BR-3, SK-BR-3, K562, A549 and A549 cells (American Type Culture Collection, Rockville, MD), respectively, using the specific primers in Table .

    Techniques: Expressing, Quantitative RT-PCR, Marker

    LunX mRNA is the most specific gene marker with high sensitivity for NSCLC cells in pleural fluid . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each pleural fluid sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as 50%, the median cannot be shown. The Mann-Whitney U Test was used to compare the gene expression levels between NSCLC and tuberculo pleurisy groups. P " width="100%" height="100%">

    Journal: BMC Cancer

    Article Title: Diagnostic utility of LunX mRNA in peripheral blood and pleural fluid in patients with primary non-small cell lung cancer

    doi: 10.1186/1471-2407-8-156

    Figure Lengend Snippet: LunX mRNA is the most specific gene marker with high sensitivity for NSCLC cells in pleural fluid . LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin mRNA from each pleural fluid sample were detected by real-time RT-PCR, and mRNA copy number was determined by reference to the standard curve, as described in methods. The copy number of each mRNA was further normalized as the ratio to the copy number of β-actin . For each gene marker, when the copy number was less than 100, it could not be detectable as a negative case. All the negative results of each indicated gene were shown as number undetected. The median is marked as " --- " in each group. Where the frequency of negative cases is > 50%, the median cannot be shown. The Mann-Whitney U Test was used to compare the gene expression levels between NSCLC and tuberculo pleurisy groups. P

    Article Snippet: Preparation of standard curves LunX, CK19, CEA, VEGF-C, hnRNP A2/B1 and β-actin cDNA were generated from A549, SK-BR-3, SK-BR-3, K562, A549 and A549 cells (American Type Culture Collection, Rockville, MD), respectively, using the specific primers in Table .

    Techniques: Marker, Quantitative RT-PCR, MANN-WHITNEY, Expressing

    Neutralizing antibody response following RSV-G protein, FI-RSV and live RSV experimental infection. ( A ) Schematic representation of cotton rat immunization and challenge schedule. Inbred female  Sigmodon hispidus  cotton rats between 6 and 8 weeks of age were immunized i.m. with PBS (Gps A-B), 5 μg of unadjuvanted or Emulsigen-adjuvanted REG ( R ecombinant  E . coli  produced  G ) (Gps C-D), or with FI-RSV (Gp E) on days 0 and 28 in groups A thru E, or were infected intranasally (i.n.) with 0.1 ml of RSV/A2 at 10 5  pfu per rat (Gp F). Blood was collected by eye-bleed on days 0, 28 and 49. On day 49, animals were either mock challenge intranasally with 0.1 ml of PBS (Gp A), or with 0.1 ml of RSV-A2 virus at 10 5  pfu per animal (10 animals per group) (Gps B-F). Cotton rats were sacrificed on days 2 or 5 post-challenge wherein lungs and nose tissues were collected. ( B ) Sera from individual cotton rats collected at pre-vaccination (day 0) and 3 weeks post second immunization (day 49) were tested for neutralization in a plaque reduction neutralization test (PRNT) against the homologous RSV-A2 strain and heterologous RSV-B1 strain. Neutralizing antibody titers represent 50% inhibition of plaque numbers. Statistical significance was tested with one-way ANOVA and Bonferroni multiple comparisons tests. ***p 

    Journal: Scientific Reports

    Article Title: Preclinical evaluation of bacterially produced RSV-G protein vaccine: Strong protection against RSV challenge in cotton rat model

    doi: 10.1038/srep42428

    Figure Lengend Snippet: Neutralizing antibody response following RSV-G protein, FI-RSV and live RSV experimental infection. ( A ) Schematic representation of cotton rat immunization and challenge schedule. Inbred female Sigmodon hispidus cotton rats between 6 and 8 weeks of age were immunized i.m. with PBS (Gps A-B), 5 μg of unadjuvanted or Emulsigen-adjuvanted REG ( R ecombinant E . coli produced G ) (Gps C-D), or with FI-RSV (Gp E) on days 0 and 28 in groups A thru E, or were infected intranasally (i.n.) with 0.1 ml of RSV/A2 at 10 5 pfu per rat (Gp F). Blood was collected by eye-bleed on days 0, 28 and 49. On day 49, animals were either mock challenge intranasally with 0.1 ml of PBS (Gp A), or with 0.1 ml of RSV-A2 virus at 10 5 pfu per animal (10 animals per group) (Gps B-F). Cotton rats were sacrificed on days 2 or 5 post-challenge wherein lungs and nose tissues were collected. ( B ) Sera from individual cotton rats collected at pre-vaccination (day 0) and 3 weeks post second immunization (day 49) were tested for neutralization in a plaque reduction neutralization test (PRNT) against the homologous RSV-A2 strain and heterologous RSV-B1 strain. Neutralizing antibody titers represent 50% inhibition of plaque numbers. Statistical significance was tested with one-way ANOVA and Bonferroni multiple comparisons tests. ***p 

    Article Snippet: The prototype A2 strain of RSV (ATCC, Manassas, VA) for challenge of cotton rats was propagated in HEp-2 cells and subjected to serial plaque-purification to reduce defective-interfering particles.

    Techniques: Infection, Produced, Neutralization, Plaque Reduction Neutralization Test, Inhibition

    GtmE Acts as an UDP-Xylose Glycosyltransferase to Generate Gentamicin A2 (4a).

    Journal:

    Article Title: Genetic dissection of the biosynthetic route to gentamicin A2 by heterologous expression of its minimal gene set

    doi: 10.1073/pnas.0803164105

    Figure Lengend Snippet: GtmE Acts as an UDP-Xylose Glycosyltransferase to Generate Gentamicin A2 (4a).

    Article Snippet: To obtain authentic gentamicin A2 , M. echinospora ATCC 15385 was grown for 7 days at 28°C in 50 ml of N-Z amine medium (1% glucose, 2% soluble starch, 0.5% yeast extract, 0.5% N-Z amine and 0.2% calcium carbonate) in 500-ml baffled flasks on a rotary shaker ( ).

    Techniques:

    HPLC-ESI-MS/MS chromatograms for the glycosyltransferase-catalyzed production of 2′- N -acetylparomamine ( 2a ) and gentamicin A2 ( 4a ). ( A–C ) GtmG assay using cell-free extracts from S. venezuelae strain harboring pYJ498 ( A ) without exogenous

    Journal:

    Article Title: Genetic dissection of the biosynthetic route to gentamicin A2 by heterologous expression of its minimal gene set

    doi: 10.1073/pnas.0803164105

    Figure Lengend Snippet: HPLC-ESI-MS/MS chromatograms for the glycosyltransferase-catalyzed production of 2′- N -acetylparomamine ( 2a ) and gentamicin A2 ( 4a ). ( A–C ) GtmG assay using cell-free extracts from S. venezuelae strain harboring pYJ498 ( A ) without exogenous

    Article Snippet: To obtain authentic gentamicin A2 , M. echinospora ATCC 15385 was grown for 7 days at 28°C in 50 ml of N-Z amine medium (1% glucose, 2% soluble starch, 0.5% yeast extract, 0.5% N-Z amine and 0.2% calcium carbonate) in 500-ml baffled flasks on a rotary shaker ( ).

    Techniques: High Performance Liquid Chromatography, Mass Spectrometry

    GtmE Acts as an UDP-Xylose Glycosyltransferase to Generate Gentamicin A2 (4a).

    Journal:

    Article Title: Genetic dissection of the biosynthetic route to gentamicin A2 by heterologous expression of its minimal gene set

    doi: 10.1073/pnas.0803164105

    Figure Lengend Snippet: GtmE Acts as an UDP-Xylose Glycosyltransferase to Generate Gentamicin A2 (4a).

    Article Snippet: To obtain authentic gentamicin A2 , M. echinospora ATCC 15385 was grown for 7 days at 28°C in 50 ml of N-Z amine medium (1% glucose, 2% soluble starch, 0.5% yeast extract, 0.5% N-Z amine and 0.2% calcium carbonate) in 500-ml baffled flasks on a rotary shaker ( ).

    Techniques:

    PCR fingerprinting profile of plasmids pMMA2 and pMCU3, determined by using a set of oligonucleotides. Lane 1, PCR fingerprint of plasmids isolated from strain AbH12O-A2; lane 2, PCR fingerprint of plasmids isolated from strain ATCC 17978 transformed

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Horizontal Transfer of the OXA-24 Carbapenemase Gene via Outer Membrane Vesicles: a New Mechanism of Dissemination of Carbapenem Resistance Genes in Acinetobacter baumannii ▿

    doi: 10.1128/AAC.00929-10

    Figure Lengend Snippet: PCR fingerprinting profile of plasmids pMMA2 and pMCU3, determined by using a set of oligonucleotides. Lane 1, PCR fingerprint of plasmids isolated from strain AbH12O-A2; lane 2, PCR fingerprint of plasmids isolated from strain ATCC 17978 transformed

    Article Snippet: In the case of clinical strains AbH12O-A2 and AbH12O-CU3, the ATCC isolate transformed with OMVs carrying clinical plasmids pMMA2 and pMMCU3 released OMVs harboring the bla OXA-24 gene ( B).

    Techniques: Polymerase Chain Reaction, Isolation, Transformation Assay

    REP-PCR profile of the A. baumannii strains used in this study. Blot A, AbH12O-A2; blot B, AbH12O-CU3; blot C, ATCC 17978; blot D, ATCC 17978 transformed with OMVs from AbH12O-A2; blot E, ATCC 17978 transformed with OMVs from AbH12O-CU3.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Horizontal Transfer of the OXA-24 Carbapenemase Gene via Outer Membrane Vesicles: a New Mechanism of Dissemination of Carbapenem Resistance Genes in Acinetobacter baumannii ▿

    doi: 10.1128/AAC.00929-10

    Figure Lengend Snippet: REP-PCR profile of the A. baumannii strains used in this study. Blot A, AbH12O-A2; blot B, AbH12O-CU3; blot C, ATCC 17978; blot D, ATCC 17978 transformed with OMVs from AbH12O-A2; blot E, ATCC 17978 transformed with OMVs from AbH12O-CU3.

    Article Snippet: In the case of clinical strains AbH12O-A2 and AbH12O-CU3, the ATCC isolate transformed with OMVs carrying clinical plasmids pMMA2 and pMMCU3 released OMVs harboring the bla OXA-24 gene ( B).

    Techniques: Polymerase Chain Reaction, Transformation Assay

    Electron microscopy micrograph of OMVs released by A. baumannii clinical strain AbH12O-A2. Vesicles were purified from broth cultures by ultracentrifugation and filtered through a 0.22-μm filter. The average diameter of the vesicles was 40 nm.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Horizontal Transfer of the OXA-24 Carbapenemase Gene via Outer Membrane Vesicles: a New Mechanism of Dissemination of Carbapenem Resistance Genes in Acinetobacter baumannii ▿

    doi: 10.1128/AAC.00929-10

    Figure Lengend Snippet: Electron microscopy micrograph of OMVs released by A. baumannii clinical strain AbH12O-A2. Vesicles were purified from broth cultures by ultracentrifugation and filtered through a 0.22-μm filter. The average diameter of the vesicles was 40 nm.

    Article Snippet: In the case of clinical strains AbH12O-A2 and AbH12O-CU3, the ATCC isolate transformed with OMVs carrying clinical plasmids pMMA2 and pMMCU3 released OMVs harboring the bla OXA-24 gene ( B).

    Techniques: Electron Microscopy, Purification

    (A) Dose-response experiment. Shown is a dot blot analysis for detecting the presence of the bla OXA- 24 gene in OMVs released from A. baumannii clinical isolate AbH12O-A2. Blot A, 10 μl of purified plasmid pMMA2 as a positive control; blot B, 10

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Horizontal Transfer of the OXA-24 Carbapenemase Gene via Outer Membrane Vesicles: a New Mechanism of Dissemination of Carbapenem Resistance Genes in Acinetobacter baumannii ▿

    doi: 10.1128/AAC.00929-10

    Figure Lengend Snippet: (A) Dose-response experiment. Shown is a dot blot analysis for detecting the presence of the bla OXA- 24 gene in OMVs released from A. baumannii clinical isolate AbH12O-A2. Blot A, 10 μl of purified plasmid pMMA2 as a positive control; blot B, 10

    Article Snippet: In the case of clinical strains AbH12O-A2 and AbH12O-CU3, the ATCC isolate transformed with OMVs carrying clinical plasmids pMMA2 and pMMCU3 released OMVs harboring the bla OXA-24 gene ( B).

    Techniques: Dot Blot, Purification, Plasmid Preparation, Positive Control