Structured Review

Millipore paa
Surface morphologies of the microspheres of (A) neat PLLA, (B) PLLA- g <t>-PAA,</t> (C) PAM g , (D) PAH2.5 g , (E) PAH5 g , (F) PAM g GO, (G) <t>PAH</t> g GO, (H) PAM g FeO, and (I) PAH g FeO.
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Images

1) Product Images from "Highlighting the Importance of Surface Grafting in Combination with a Layer-by-Layer Approach for Fabricating Advanced 3D Poly(l-lactide) Microsphere Scaffolds"

Article Title: Highlighting the Importance of Surface Grafting in Combination with a Layer-by-Layer Approach for Fabricating Advanced 3D Poly(l-lactide) Microsphere Scaffolds

Journal: Chemistry of Materials

doi: 10.1021/acs.chemmater.6b00133

Surface morphologies of the microspheres of (A) neat PLLA, (B) PLLA- g -PAA, (C) PAM g , (D) PAH2.5 g , (E) PAH5 g , (F) PAM g GO, (G) PAH g GO, (H) PAM g FeO, and (I) PAH g FeO.
Figure Legend Snippet: Surface morphologies of the microspheres of (A) neat PLLA, (B) PLLA- g -PAA, (C) PAM g , (D) PAH2.5 g , (E) PAH5 g , (F) PAM g GO, (G) PAH g GO, (H) PAM g FeO, and (I) PAH g FeO.

Techniques Used:

ATR-FTIR spectra of neat PLLA microspheres (black) and PEMs alternately assembled onto the surface of PLLA- g -PAA microspheres with PAM g (red), PAM g GO (magenta), PAM g FeO (orange), PAH2.5 g (green), PAH g GO (violet), PAH g FeO (gray), and PAH5 g (blue) in the region of (A) 1680–1500 cm –1 and (B) 3800–2300 cm –1 . (C) XRD patterns of the 3D microsphere scaffolds with and without GO functionalization.
Figure Legend Snippet: ATR-FTIR spectra of neat PLLA microspheres (black) and PEMs alternately assembled onto the surface of PLLA- g -PAA microspheres with PAM g (red), PAM g GO (magenta), PAM g FeO (orange), PAH2.5 g (green), PAH g GO (violet), PAH g FeO (gray), and PAH5 g (blue) in the region of (A) 1680–1500 cm –1 and (B) 3800–2300 cm –1 . (C) XRD patterns of the 3D microsphere scaffolds with and without GO functionalization.

Techniques Used:

2) Product Images from "A potent novel vaccine adjuvant based on straight polyacrylate"

Article Title: A potent novel vaccine adjuvant based on straight polyacrylate

Journal: International Journal of Pharmaceutics: X

doi: 10.1016/j.ijpx.2020.100054

Adjuvant activity of different technical lots of PAA is dependent on polymer size and dose and not on polymer branching. C57BL/6J mice (n = 10/group) were immunized 2 times, 25 days apart, by IM injection of 2 μg of recombinant CMV-gB without adjuvant or with MF59 or different PAAs at increasing concentrations (25-50-100-200-400 μg/dose) under a final volume of 50 μl. The PAAs used in this experiment were lots # 566253 (○), #652979 (□), #240413 (∆) and #655555 (◊) characterized in Table 1 . Two weeks following the second administration (D39), CMV neutralizing titers determined on ARPE-19 cells (Panel A), CMV-gB-specific IgG1 (panel B) and IgG2c (panel C) titers and spleen cell IFN-γ responses (panel D) were determined as in Fig. 1 . Antibody and IFN-γ levels obtained in mice injected with CMV-gB alone or combined with MF59, are shown as dotted and dash lines, respectively. No IL-5 was detected in the PAA groups in this study.
Figure Legend Snippet: Adjuvant activity of different technical lots of PAA is dependent on polymer size and dose and not on polymer branching. C57BL/6J mice (n = 10/group) were immunized 2 times, 25 days apart, by IM injection of 2 μg of recombinant CMV-gB without adjuvant or with MF59 or different PAAs at increasing concentrations (25-50-100-200-400 μg/dose) under a final volume of 50 μl. The PAAs used in this experiment were lots # 566253 (○), #652979 (□), #240413 (∆) and #655555 (◊) characterized in Table 1 . Two weeks following the second administration (D39), CMV neutralizing titers determined on ARPE-19 cells (Panel A), CMV-gB-specific IgG1 (panel B) and IgG2c (panel C) titers and spleen cell IFN-γ responses (panel D) were determined as in Fig. 1 . Antibody and IFN-γ levels obtained in mice injected with CMV-gB alone or combined with MF59, are shown as dotted and dash lines, respectively. No IL-5 was detected in the PAA groups in this study.

Techniques Used: Activity Assay, Mouse Assay, Injection, Recombinant

Adjuvant activity of PAA is strongly dependent on polymer size. C57BL/6J mice ( n = 10/group) were immunized 2 times, 4 weeks apart, by IM injection of 2 μg of recombinant CMV-gB without adjuvant (white bars) or with MF59 (red bars) or PAA (blue bars) under a final volume of 50 μl. Low Mw PAA (lot#617193; Mw = 9 kDa and PI = 3.1), medium Mw PAA (lot#601318; Mw = 134 kDa and PI = 2.4) and high molecular weight PAA (lot#566253; Mw = 489 kDa and PI = 3.8) were all tested at 200 μg/dose. Two weeks after the second immunization (D42), specific serum antibody responses and spleen cell cytokine responses were determined as described in the Materials and Methods section. Panel A: CMV neutralizing antibody titers measured on ARPE-19 epithelial cells. Panel B: CMV neutralizing antibody titers measured on MRC-5 fibroblasts. Panel C: CMV-gB-specific IgG1 and IgG2c titers. Panel D: IL-5 and IFN-γ productions from restimulated splenocytes cultures. Titers were plotted as geometric mean titers with 95% Confidence Interval (CI) on a log scale. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
Figure Legend Snippet: Adjuvant activity of PAA is strongly dependent on polymer size. C57BL/6J mice ( n = 10/group) were immunized 2 times, 4 weeks apart, by IM injection of 2 μg of recombinant CMV-gB without adjuvant (white bars) or with MF59 (red bars) or PAA (blue bars) under a final volume of 50 μl. Low Mw PAA (lot#617193; Mw = 9 kDa and PI = 3.1), medium Mw PAA (lot#601318; Mw = 134 kDa and PI = 2.4) and high molecular weight PAA (lot#566253; Mw = 489 kDa and PI = 3.8) were all tested at 200 μg/dose. Two weeks after the second immunization (D42), specific serum antibody responses and spleen cell cytokine responses were determined as described in the Materials and Methods section. Panel A: CMV neutralizing antibody titers measured on ARPE-19 epithelial cells. Panel B: CMV neutralizing antibody titers measured on MRC-5 fibroblasts. Panel C: CMV-gB-specific IgG1 and IgG2c titers. Panel D: IL-5 and IFN-γ productions from restimulated splenocytes cultures. Titers were plotted as geometric mean titers with 95% Confidence Interval (CI) on a log scale. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Techniques Used: Activity Assay, Mouse Assay, Injection, Recombinant, Molecular Weight

3) Product Images from "Stable and non-toxic ultrasmall gadolinium oxide nanoparticle colloids (coating material = polyacrylic acid) as high-performance T1 magnetic resonance imaging contrast agents"

Article Title: Stable and non-toxic ultrasmall gadolinium oxide nanoparticle colloids (coating material = polyacrylic acid) as high-performance T1 magnetic resonance imaging contrast agents

Journal: RSC Advances

doi: 10.1039/c7ra11830a

TGA curve showing the PAA surface-coating amount (53.7%) in weight percent. The weight percents of water and air (18.4%) and Gd 2 O 3 (27.9%) are also provided.
Figure Legend Snippet: TGA curve showing the PAA surface-coating amount (53.7%) in weight percent. The weight percents of water and air (18.4%) and Gd 2 O 3 (27.9%) are also provided.

Techniques Used:

Reaction scheme for the one-pot synthesis of ultrasmall Gd 2 O 3 nanoparticle colloid and PAA structure.
Figure Legend Snippet: Reaction scheme for the one-pot synthesis of ultrasmall Gd 2 O 3 nanoparticle colloid and PAA structure.

Techniques Used:

(a) FT-IR absorption spectra of a powder sample and free PAA ( M w = ∼5100 Da): 3325 cm −1 (O–H stretch from water); 2930 cm −1 (C–H stretch from PAA); 1660 cm −1 (O–H bend from water); 1550 cm −1 (COO − antisymmetric stretch from PAA); and 1400 cm −1 (COO − symmetric stretch from PAA). (b) Surface-coating structure of PAA on the ultrasmall Gd 2 O 3 nanoparticle surface.
Figure Legend Snippet: (a) FT-IR absorption spectra of a powder sample and free PAA ( M w = ∼5100 Da): 3325 cm −1 (O–H stretch from water); 2930 cm −1 (C–H stretch from PAA); 1660 cm −1 (O–H bend from water); 1550 cm −1 (COO − antisymmetric stretch from PAA); and 1400 cm −1 (COO − symmetric stretch from PAA). (b) Surface-coating structure of PAA on the ultrasmall Gd 2 O 3 nanoparticle surface.

Techniques Used:

4) Product Images from "Varicella-Zoster Virus Retains Major Histocompatibility Complex Class I Proteins in the Golgi Compartment of Infected Cells"

Article Title: Varicella-Zoster Virus Retains Major Histocompatibility Complex Class I Proteins in the Golgi Compartment of Infected Cells

Journal: Journal of Virology

doi: 10.1128/JVI.75.10.4878-4888.2001

Analysis of MHC I downregulation in VZV- and mock-infected cells treated with the viral DNA inhibitor PAA. (A) Western blot of viral gC was performed on total cell lysates from mock-infected cells (lane 1), mock-infected cells with PAA (lane 2), VZV-infected cells at the time of inoculation onto uninfected cells (lane 3), and VZV-infected cells in the absence (lane 4) or presence (lane 5) of PAA at 24 h postinfection. (B) The percentage of VZV + and VZV − cell populations expressing cell surface MHC I molecules. Cells were infected with VZV in the absence or presence of PAA for 24 h and stained with antibodies and fluorescent conjugates to MHC I and VZV proteins and analyzed by flow cytometry.
Figure Legend Snippet: Analysis of MHC I downregulation in VZV- and mock-infected cells treated with the viral DNA inhibitor PAA. (A) Western blot of viral gC was performed on total cell lysates from mock-infected cells (lane 1), mock-infected cells with PAA (lane 2), VZV-infected cells at the time of inoculation onto uninfected cells (lane 3), and VZV-infected cells in the absence (lane 4) or presence (lane 5) of PAA at 24 h postinfection. (B) The percentage of VZV + and VZV − cell populations expressing cell surface MHC I molecules. Cells were infected with VZV in the absence or presence of PAA for 24 h and stained with antibodies and fluorescent conjugates to MHC I and VZV proteins and analyzed by flow cytometry.

Techniques Used: Infection, Western Blot, Expressing, Staining, Flow Cytometry, Cytometry

5) Product Images from "Effective Inhibition of Kb- and Db-Restricted Antigen Presentation in Primary Macrophages by Murine Cytomegalovirus"

Article Title: Effective Inhibition of Kb- and Db-Restricted Antigen Presentation in Primary Macrophages by Murine Cytomegalovirus

Journal: Journal of Virology

doi: 10.1128/JVI.77.1.301-308.2003

Isolation and infection of BMMφ. (A) Isolation of a pure population of macrophages. Bone marrow was flushed from the femurs and cultured for 6 days on non-tissue-culture-treated plastic petri dishes with L929 supernatant as a source of GM-CSF. Adherent cells were removed with cold PBS, stained with anti-F4/80 or isotype control, and analyzed by FACS. (B) BMMφ and IFN-γ-pretreated MEFs were plated onto glass coverslips and infected overnight with wild-type MCMV (MW97.01) in the presence of PAA at the indicated MOIs. Cells were stained with rabbit antiserum recognizing m4/gp34 or with NRS and were treated with Hoescht DNA stain to detect all cells. Percent cells expressing m4 was determined by the ratio of cells expressing m4/gp34 to Hoechst-staining cells in the same fields. (C) BMMφ were treated with IFN-γ (50 U/ml) for 24 h or left untreated and then infected overnight as described above at an MOI of 50. Percent cells expressing m4 was determined as described above.
Figure Legend Snippet: Isolation and infection of BMMφ. (A) Isolation of a pure population of macrophages. Bone marrow was flushed from the femurs and cultured for 6 days on non-tissue-culture-treated plastic petri dishes with L929 supernatant as a source of GM-CSF. Adherent cells were removed with cold PBS, stained with anti-F4/80 or isotype control, and analyzed by FACS. (B) BMMφ and IFN-γ-pretreated MEFs were plated onto glass coverslips and infected overnight with wild-type MCMV (MW97.01) in the presence of PAA at the indicated MOIs. Cells were stained with rabbit antiserum recognizing m4/gp34 or with NRS and were treated with Hoescht DNA stain to detect all cells. Percent cells expressing m4 was determined by the ratio of cells expressing m4/gp34 to Hoechst-staining cells in the same fields. (C) BMMφ were treated with IFN-γ (50 U/ml) for 24 h or left untreated and then infected overnight as described above at an MOI of 50. Percent cells expressing m4 was determined as described above.

Techniques Used: Isolation, Infection, Cell Culture, Staining, FACS, Expressing

m152/gp40 prevents CTL recognition in IC21 macrophages. MEFs and IC21 macrophages were treated with IFN-γ and infected at an MOI of 70 in the presence of PAA with wild-type MCMV (MW97.01) or Δm152 (MW99.05). They were then used as targets in a Cr release assay with the D b -restricted, M45-specific CTL clone 3. Error bars represent the standard errors of the means. E:T, effector-to-target ratio; SL, specific lysis.
Figure Legend Snippet: m152/gp40 prevents CTL recognition in IC21 macrophages. MEFs and IC21 macrophages were treated with IFN-γ and infected at an MOI of 70 in the presence of PAA with wild-type MCMV (MW97.01) or Δm152 (MW99.05). They were then used as targets in a Cr release assay with the D b -restricted, M45-specific CTL clone 3. Error bars represent the standard errors of the means. E:T, effector-to-target ratio; SL, specific lysis.

Techniques Used: CTL Assay, Infection, Release Assay, Lysis

m4/gp34 prevents antigen presentation in BMMφ. BMMφ and IFN-γ-pretreated MEF targets were 51 Cr loaded and infected at an MOI of 100 overnight, in the presence of PAA, with no virus, wild-type MCMV (MW97.01), Δm4 (MW99.03), or Δm152 (MW99.05). K b -restricted CTL clones 5, 11, and 96 and D b -restricted clones 3 and 55 were tested for their ability to recognize and lyse targets. E:T, effector-to-target ratio; SL, specific lysis.
Figure Legend Snippet: m4/gp34 prevents antigen presentation in BMMφ. BMMφ and IFN-γ-pretreated MEF targets were 51 Cr loaded and infected at an MOI of 100 overnight, in the presence of PAA, with no virus, wild-type MCMV (MW97.01), Δm4 (MW99.03), or Δm152 (MW99.05). K b -restricted CTL clones 5, 11, and 96 and D b -restricted clones 3 and 55 were tested for their ability to recognize and lyse targets. E:T, effector-to-target ratio; SL, specific lysis.

Techniques Used: Infection, CTL Assay, Clone Assay, Lysis

m152/gp40 retains class I MHC molecules and interferes with antigen presentation in MEFs and BMMφ. (A) m152 retains MHC class I molecules in a pre-Golgi compartment in BMMφ. MEFs were pretreated with IFN-γ for 48 h. MEFs and BMMφ were infected with wild-type MCMV (Smith), Δm152 (MC96.24), or revertant Δm152 (rMC96.27) or were left uninfected. Cells were 35 S labeled overnight in the presence of 0.3 mg of PAA/ml. Cell lysates were subjected to immunoprecipitation (IP), endo H treated, and run on an SDS-12.5% polyacrylamide gel electrophoresis gel. MEF lysates were immunoprecipitated with serum 8010 (rabbit anti-K b ), NRS control for 8010, 28.14.8S (anti-D b ), or normal mouse serum (NMS) control for 28.14.8S. endo H-resistant (R) and -sensitive (S) forms of K b , D b ) and on the appearance of these bands in cells infected with virus lacking m6 (data not shown). (B) m152 inhibits CTL recognition of MCMV-infected BMMφ. BMMφ and IFN-γ-pretreated MEF targets were 51 Cr loaded and infected at an MOI of 45 overnight, in the presence of PAA, with no virus, Smith (wild-type MCMV), Δm152 (MC96.24), or revertant Δm152 (rMC96.27). K b -restricted CTL clone 5 and D b -restricted clone 3 were tested for the ability to recognize and lyse targets. SL, specific lysis. (C) Wild-type virus is poorly recognized in BMMφ regardless of MOI. Targets were infected as described before with Δm152 (MW99.05) or wild-type (MW97.01) virus at the MOIs indicated and were tested for lysis by CTL clones 5 and 96 (both K b restricted). The effector-to-target ratio (E:T) for both clones was 20:1 for MEFs and 5:1 for BMMφ. (D) PAA does not affect recognition of wild-type virus in BMMφ. Targets were infected as described before in the presence or absence of PAA, and a CTL assay was performed as before using a mixture of clones 3 and 55 (both specific for M45 restricted by D b ).
Figure Legend Snippet: m152/gp40 retains class I MHC molecules and interferes with antigen presentation in MEFs and BMMφ. (A) m152 retains MHC class I molecules in a pre-Golgi compartment in BMMφ. MEFs were pretreated with IFN-γ for 48 h. MEFs and BMMφ were infected with wild-type MCMV (Smith), Δm152 (MC96.24), or revertant Δm152 (rMC96.27) or were left uninfected. Cells were 35 S labeled overnight in the presence of 0.3 mg of PAA/ml. Cell lysates were subjected to immunoprecipitation (IP), endo H treated, and run on an SDS-12.5% polyacrylamide gel electrophoresis gel. MEF lysates were immunoprecipitated with serum 8010 (rabbit anti-K b ), NRS control for 8010, 28.14.8S (anti-D b ), or normal mouse serum (NMS) control for 28.14.8S. endo H-resistant (R) and -sensitive (S) forms of K b , D b ) and on the appearance of these bands in cells infected with virus lacking m6 (data not shown). (B) m152 inhibits CTL recognition of MCMV-infected BMMφ. BMMφ and IFN-γ-pretreated MEF targets were 51 Cr loaded and infected at an MOI of 45 overnight, in the presence of PAA, with no virus, Smith (wild-type MCMV), Δm152 (MC96.24), or revertant Δm152 (rMC96.27). K b -restricted CTL clone 5 and D b -restricted clone 3 were tested for the ability to recognize and lyse targets. SL, specific lysis. (C) Wild-type virus is poorly recognized in BMMφ regardless of MOI. Targets were infected as described before with Δm152 (MW99.05) or wild-type (MW97.01) virus at the MOIs indicated and were tested for lysis by CTL clones 5 and 96 (both K b restricted). The effector-to-target ratio (E:T) for both clones was 20:1 for MEFs and 5:1 for BMMφ. (D) PAA does not affect recognition of wild-type virus in BMMφ. Targets were infected as described before in the presence or absence of PAA, and a CTL assay was performed as before using a mixture of clones 3 and 55 (both specific for M45 restricted by D b ).

Techniques Used: Infection, Labeling, Immunoprecipitation, Polyacrylamide Gel Electrophoresis, CTL Assay, Lysis, Clone Assay

6) Product Images from "Ferroferric oxide nanoparticles induce prosurvival autophagy in human blood cells by modulating the Beclin 1/Bcl-2/VPS34 complex"

Article Title: Ferroferric oxide nanoparticles induce prosurvival autophagy in human blood cells by modulating the Beclin 1/Bcl-2/VPS34 complex

Journal: International Journal of Nanomedicine

doi: 10.2147/IJN.S72598

Characterization of various surface modifications of superparamagnetic nanoparticles. Notes:  TEM images of ( A ) Fe 3 O 4 -DA, ( B ) Fe 3 O 4 -DMSA, and ( C ) Fe 3 O 4 -PEG. ( D ) DLS size of Fe 3 O 4 -DA, Fe 3 O 4 -DMSA, and Fe 3 O 4 -PEG in water solution; ( E ) Zeta potential of the Fe 3 O 4 -DA, Fe 3 O 4 -DMSA, and Fe 3 O 4 -PEG in water solution. Photos of ( F ) Fe 3 O 4 -DA, ( G ) Fe 3 O 4 -DMSA, and ( H ) Fe 3 O 4 -PEG in various solutions: water, phosphate buffered saline (PBS), RMPI-1640 cell medium, and fetal bovine serum (FBS) from left to right. Abbreviations:  DA, dopamine; DMSA, dimercaptosuccinic acid; DLS, dynamic light scattering; Fe 3 O 4 , ferroferric oxide; PEG, polyethylene glycol; TEM, transmission electronic microscopy.
Figure Legend Snippet: Characterization of various surface modifications of superparamagnetic nanoparticles. Notes: TEM images of ( A ) Fe 3 O 4 -DA, ( B ) Fe 3 O 4 -DMSA, and ( C ) Fe 3 O 4 -PEG. ( D ) DLS size of Fe 3 O 4 -DA, Fe 3 O 4 -DMSA, and Fe 3 O 4 -PEG in water solution; ( E ) Zeta potential of the Fe 3 O 4 -DA, Fe 3 O 4 -DMSA, and Fe 3 O 4 -PEG in water solution. Photos of ( F ) Fe 3 O 4 -DA, ( G ) Fe 3 O 4 -DMSA, and ( H ) Fe 3 O 4 -PEG in various solutions: water, phosphate buffered saline (PBS), RMPI-1640 cell medium, and fetal bovine serum (FBS) from left to right. Abbreviations: DA, dopamine; DMSA, dimercaptosuccinic acid; DLS, dynamic light scattering; Fe 3 O 4 , ferroferric oxide; PEG, polyethylene glycol; TEM, transmission electronic microscopy.

Techniques Used: Transmission Electron Microscopy, Transmission Assay, Microscopy

7) Product Images from "ATRX promotes maintenance of herpes simplex virus heterochromatin during chromatin stress"

Article Title: ATRX promotes maintenance of herpes simplex virus heterochromatin during chromatin stress

Journal: eLife

doi: 10.7554/eLife.40228

ATRX restricts HSV gene expression from input and progeny viral DNA. ( A ) HFFs were infected with HSV 7134 at an MOI of 3, and infected cells were fixed and harvested 30, 60, and 240 min post infection. ChIP-qCPR and HSV specific primers were used to detect chromatin enrichment of ATRX at ICP27 (blue) and ICP8 (black) gene promoters. Two-tailed t-tests were used to compare ATRX enrichment at viral gene promoters compared to GAPDH. ( B ) HFFs were treated with siNT or siATRX and infected with HSV 7134 at an MOI of 5 in the absence (left panels) or presence (right panels) of PAA. Relative viral transcripts for ( B ) ICP27 , ( C ) ICP8 , or ( D ) gB were quantified by qPCR at 0, 2, 4, 6, and 8 hpi. Viral mRNA levels were normalized to cellular 18S transcripts. Results were analyzed by two-way ANOVA. All data for Figure 3 are reported as the average of 3 independent experiments ± standard error of the mean; p
Figure Legend Snippet: ATRX restricts HSV gene expression from input and progeny viral DNA. ( A ) HFFs were infected with HSV 7134 at an MOI of 3, and infected cells were fixed and harvested 30, 60, and 240 min post infection. ChIP-qCPR and HSV specific primers were used to detect chromatin enrichment of ATRX at ICP27 (blue) and ICP8 (black) gene promoters. Two-tailed t-tests were used to compare ATRX enrichment at viral gene promoters compared to GAPDH. ( B ) HFFs were treated with siNT or siATRX and infected with HSV 7134 at an MOI of 5 in the absence (left panels) or presence (right panels) of PAA. Relative viral transcripts for ( B ) ICP27 , ( C ) ICP8 , or ( D ) gB were quantified by qPCR at 0, 2, 4, 6, and 8 hpi. Viral mRNA levels were normalized to cellular 18S transcripts. Results were analyzed by two-way ANOVA. All data for Figure 3 are reported as the average of 3 independent experiments ± standard error of the mean; p

Techniques Used: Expressing, Infection, Chromatin Immunoprecipitation, Two Tailed Test, Real-time Polymerase Chain Reaction

EdC labeling of KOS HSV. ( A ) HFF cells infected with HSV-EdC at an MOI of 5 and fixed at 2 hpi. vDNA is shown in green and viral protein ICP4 in red. ( B ) HFF cells infected with HSV-EdC at an MOI of 5 in the absence (-) or presence (+) of viral DNA synthesis inhibitor PAA and fixed at 4 and 6 hpi. vDNA is shown in green and viral protein ICP8 in red. ( C ) Immunoblot comparing ICP4 and ICP8 from HSV and HSV-EdC at 4, 8, and 12 hpi. ( D ) HFF cells infected with HSV-EdC at an MOI of 5 in the absence (-) or presence (+) heparin at 1 hpi. HSV-EdC genomes in green.
Figure Legend Snippet: EdC labeling of KOS HSV. ( A ) HFF cells infected with HSV-EdC at an MOI of 5 and fixed at 2 hpi. vDNA is shown in green and viral protein ICP4 in red. ( B ) HFF cells infected with HSV-EdC at an MOI of 5 in the absence (-) or presence (+) of viral DNA synthesis inhibitor PAA and fixed at 4 and 6 hpi. vDNA is shown in green and viral protein ICP8 in red. ( C ) Immunoblot comparing ICP4 and ICP8 from HSV and HSV-EdC at 4, 8, and 12 hpi. ( D ) HFF cells infected with HSV-EdC at an MOI of 5 in the absence (-) or presence (+) heparin at 1 hpi. HSV-EdC genomes in green.

Techniques Used: Labeling, Infection, DNA Synthesis

8) Product Images from "Breathing silicon anodes for durable high-power operations"

Article Title: Breathing silicon anodes for durable high-power operations

Journal: Scientific Reports

doi: 10.1038/srep14433

Mechanical properties and related molecular configuration. ( a ) Hardness and elastic modulus of three different binder systems characterized by nanoindentation: PAA, PAA/CMC and PAA/pullulan. ( b ) Macroscopic crack development on silicon-based electrodes after folding and crumpling. The electrodes were made of npSi, a binder and carbon black in 6:2:2 weight ratio (loading density = ~0.8 mg cm −2 ). PAA/pullulan or PAA/CMC was used as the binder. ( c ) Molecular-level conformational changes of pyranose units of different glycosidic linkages during elongation. β-(1 → 4) represents CMC while α-(1 → 4) and α-(1 → 6) represent pullulan. Chair-to-boat conformation was found in α cases while the conformation was fixed at chair for β case even after elongation. ( d ) Simulated stress-strain curves for PVDF (120 of 25 mers), pullulan (30 of 10 mers) and CMC (30 of 30 mers). The maximum strain represents double elongation of original systems. ( e ) Chair-to-boat conformational changes in polymer system level. Pullulan (30 of 10 mers) and CMC (30 of 30 mers) were doubly elongated in one direction. The initial and final states were presented on the left and right, respectively. A polymer chain was distinguished from another by a different scale of grey. The pyranose in boat conformation was indicated by yellow.
Figure Legend Snippet: Mechanical properties and related molecular configuration. ( a ) Hardness and elastic modulus of three different binder systems characterized by nanoindentation: PAA, PAA/CMC and PAA/pullulan. ( b ) Macroscopic crack development on silicon-based electrodes after folding and crumpling. The electrodes were made of npSi, a binder and carbon black in 6:2:2 weight ratio (loading density = ~0.8 mg cm −2 ). PAA/pullulan or PAA/CMC was used as the binder. ( c ) Molecular-level conformational changes of pyranose units of different glycosidic linkages during elongation. β-(1 → 4) represents CMC while α-(1 → 4) and α-(1 → 6) represent pullulan. Chair-to-boat conformation was found in α cases while the conformation was fixed at chair for β case even after elongation. ( d ) Simulated stress-strain curves for PVDF (120 of 25 mers), pullulan (30 of 10 mers) and CMC (30 of 30 mers). The maximum strain represents double elongation of original systems. ( e ) Chair-to-boat conformational changes in polymer system level. Pullulan (30 of 10 mers) and CMC (30 of 30 mers) were doubly elongated in one direction. The initial and final states were presented on the left and right, respectively. A polymer chain was distinguished from another by a different scale of grey. The pyranose in boat conformation was indicated by yellow.

Techniques Used:

Chemical identification. ( a ) Molecular structure of three different polysaccharides: CMC, alginate and pullulan. Types of glycosidic linkages were indicated. ( b ) Covalent inter-linking between npSi and PAA as well as between PAA and pullulan after thermal treatment at 150 °C. Condensation reactions are induced at the temperature between carboxylic groups of PAA and hydroxyl groups of pullulan and surface oxide of silicon. ( c ) Infrared spectra confirming the covalent inter-linking by hydroxyl group disappearance and ester group generation.
Figure Legend Snippet: Chemical identification. ( a ) Molecular structure of three different polysaccharides: CMC, alginate and pullulan. Types of glycosidic linkages were indicated. ( b ) Covalent inter-linking between npSi and PAA as well as between PAA and pullulan after thermal treatment at 150 °C. Condensation reactions are induced at the temperature between carboxylic groups of PAA and hydroxyl groups of pullulan and surface oxide of silicon. ( c ) Infrared spectra confirming the covalent inter-linking by hydroxyl group disappearance and ester group generation.

Techniques Used:

Electrochemical characterization. ( a ) Potential profiles of a coin-type half-cell containing an electrode of silicon and carbon black with PAA/pullulan as a binder on lithiation and delithiation at representatively selected cycles. Each cycle consisted of lithiation followed by delithiation. The cell was galvanostatically lithiated at 0.2 C as well as delithiated at 0.5 C in a potential range of 0.01 V to 1.2 V. ( b ) Delithiation capacities of three different binder systems as cycle progress. The same operational condition was used as in a. ( c ) Potential profiles of a PAA/pullulan cell at different delithiation rates with a fixed lithiation rate at 0.2 C. ( d ) Delithiation capacities of PAA/pullulan and PAA/CMC-based cells at different C rates. ( e ) Capacity retention with high-rate cycles consisting of 6 C lithiation followed by 6 C delithiation without potentiostatic retention. Capacities were calculated as per silicon mass.
Figure Legend Snippet: Electrochemical characterization. ( a ) Potential profiles of a coin-type half-cell containing an electrode of silicon and carbon black with PAA/pullulan as a binder on lithiation and delithiation at representatively selected cycles. Each cycle consisted of lithiation followed by delithiation. The cell was galvanostatically lithiated at 0.2 C as well as delithiated at 0.5 C in a potential range of 0.01 V to 1.2 V. ( b ) Delithiation capacities of three different binder systems as cycle progress. The same operational condition was used as in a. ( c ) Potential profiles of a PAA/pullulan cell at different delithiation rates with a fixed lithiation rate at 0.2 C. ( d ) Delithiation capacities of PAA/pullulan and PAA/CMC-based cells at different C rates. ( e ) Capacity retention with high-rate cycles consisting of 6 C lithiation followed by 6 C delithiation without potentiostatic retention. Capacities were calculated as per silicon mass.

Techniques Used:

Morphological and dimensional changes on lithiation. ( a ) Electron-microscopic snapshots of silicon-based electrodes in top and cross-sectional views before (top) and after (bottom) lithiation. PAA/pullulan and PAA/CMC-based electrodes were compared. ( b ) 2D schematic description of morphological and dimensional changes on lithiation. Each square consists of three categorized components: npSi (blue circle), binder + carbon black (red triangle) and void (the rest part with no color). The area of each component represents its volume fraction. The connectivity among npSi, binder and carbon black was not considered. Porosity and void volume expansion coefficient (β = void volume ratios of before to after lithiation) were indicated. Also, 3D cartoon corresponding to each 2D description was demonstrated at the bottom.
Figure Legend Snippet: Morphological and dimensional changes on lithiation. ( a ) Electron-microscopic snapshots of silicon-based electrodes in top and cross-sectional views before (top) and after (bottom) lithiation. PAA/pullulan and PAA/CMC-based electrodes were compared. ( b ) 2D schematic description of morphological and dimensional changes on lithiation. Each square consists of three categorized components: npSi (blue circle), binder + carbon black (red triangle) and void (the rest part with no color). The area of each component represents its volume fraction. The connectivity among npSi, binder and carbon black was not considered. Porosity and void volume expansion coefficient (β = void volume ratios of before to after lithiation) were indicated. Also, 3D cartoon corresponding to each 2D description was demonstrated at the bottom.

Techniques Used:

9) Product Images from "Direct Repeat 6 from Human Herpesvirus-6B Encodes a Nuclear Protein that Forms a Complex with the Viral DNA Processivity Factor p41"

Article Title: Direct Repeat 6 from Human Herpesvirus-6B Encodes a Nuclear Protein that Forms a Complex with the Viral DNA Processivity Factor p41

Journal: PLoS ONE

doi: 10.1371/journal.pone.0007457

Expression of dr6 mRNA and protein in HHV-6B-infected cells. (A–D) Relative expression of dr6 mRNA using tbp mRNA as reference. HCT116 (A and B) or MOLT3 (C and D) cells were incubated in the presence or absence of CHX (A and C) or PAA (B and D) and lysed at the indicated timepoint followed by mRNA extraction. (E) Anti-DR6 polyclonal antibodies identified a 44 kDa band by Western blotting, which was blocked by the addition of a DR6(6B) peptide. The peptide sequence was as shown in Figure 1D . (F) Western blotting of HCT116 cell extract at 24, 48, and 72 hpi. Expression of DR6(6B) and GAPDH is shown. A representative of 3 experiments is shown.
Figure Legend Snippet: Expression of dr6 mRNA and protein in HHV-6B-infected cells. (A–D) Relative expression of dr6 mRNA using tbp mRNA as reference. HCT116 (A and B) or MOLT3 (C and D) cells were incubated in the presence or absence of CHX (A and C) or PAA (B and D) and lysed at the indicated timepoint followed by mRNA extraction. (E) Anti-DR6 polyclonal antibodies identified a 44 kDa band by Western blotting, which was blocked by the addition of a DR6(6B) peptide. The peptide sequence was as shown in Figure 1D . (F) Western blotting of HCT116 cell extract at 24, 48, and 72 hpi. Expression of DR6(6B) and GAPDH is shown. A representative of 3 experiments is shown.

Techniques Used: Expressing, Infection, Incubation, Western Blot, Sequencing

10) Product Images from "Adaptive growth factor delivery from a polyelectrolyte coating promotes synergistic bone tissue repair and reconstruction"

Article Title: Adaptive growth factor delivery from a polyelectrolyte coating promotes synergistic bone tissue repair and reconstruction

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1408035111

Materials used for PEM multilayer mediate bone repair. ( A ) Molecular structures of materials in the system. Hydrophobic PLGA is used to form the membrane. Poly2, PAA, BMP-2, and PDGF-BB are part of the bioactive interface that initiates the bone wound-healing
Figure Legend Snippet: Materials used for PEM multilayer mediate bone repair. ( A ) Molecular structures of materials in the system. Hydrophobic PLGA is used to form the membrane. Poly2, PAA, BMP-2, and PDGF-BB are part of the bioactive interface that initiates the bone wound-healing

Techniques Used: End-sequence Profiling

11) Product Images from "Impact of wet-dry cycling on the phase behavior and compartmentalization properties of complex coacervates"

Article Title: Impact of wet-dry cycling on the phase behavior and compartmentalization properties of complex coacervates

Journal: Nature Communications

doi: 10.1038/s41467-020-19184-z

Comparison between dried coacervates and composition mimics samples. a The solid line on the phase diagram, obtained by turbidity measurements, shows the range of the composition mimics. The dotted line shows the extension where the SAXS measurements were performed (Fig. 6 ). b Total volume of solution to coacervate phase volume ratio. The error bars in b represent a constant standard deviation of ±0.25 µL, a difference in volume that was difficult to discern with the naked eye (Supplementary Fig. 22 ). c U15 concentrations measured within the coacervate. d Partitioning coefficient of U15 ([U15] coacervate phase /[U15] dilute phase ). e Whole-droplet FRAP, and f partial-droplet FRAP in dried PDADMA/PAA coacervate samples and mimics with compositions on the same range obtained during drying. Error bars in c–f represent standard deviations among different trials and individual data points are shown in the SI (Supplementary Fig. 24 , Supplementary Tables 8 and 9 ). In c , mimics data represent means and standard deviations of three trials with 15 droplets analyzed over three images, per trial. FRAP mimics data are averages and standard deviations obtained from three trials with at least two bleached droplets per trial for whole-droplet experiments, and three bleached areas per sample for partial-droplet measurements. Photos of bromophenol blue-dyed dried and mimics samples are included above for clarification.
Figure Legend Snippet: Comparison between dried coacervates and composition mimics samples. a The solid line on the phase diagram, obtained by turbidity measurements, shows the range of the composition mimics. The dotted line shows the extension where the SAXS measurements were performed (Fig. 6 ). b Total volume of solution to coacervate phase volume ratio. The error bars in b represent a constant standard deviation of ±0.25 µL, a difference in volume that was difficult to discern with the naked eye (Supplementary Fig. 22 ). c U15 concentrations measured within the coacervate. d Partitioning coefficient of U15 ([U15] coacervate phase /[U15] dilute phase ). e Whole-droplet FRAP, and f partial-droplet FRAP in dried PDADMA/PAA coacervate samples and mimics with compositions on the same range obtained during drying. Error bars in c–f represent standard deviations among different trials and individual data points are shown in the SI (Supplementary Fig. 24 , Supplementary Tables 8 and 9 ). In c , mimics data represent means and standard deviations of three trials with 15 droplets analyzed over three images, per trial. FRAP mimics data are averages and standard deviations obtained from three trials with at least two bleached droplets per trial for whole-droplet experiments, and three bleached areas per sample for partial-droplet measurements. Photos of bromophenol blue-dyed dried and mimics samples are included above for clarification.

Techniques Used: Standard Deviation, Clarification Assay

SAXS data of complex coacervate mimics samples. a Small-angle X-ray scattering plots of PDADMA/PAA complex coacervate samples prepared at 50–100 mM polymer charge and NaCl, with HEPES ranging between 25 and 50 mM, and MgCl 2 between 4.3 and 8.6 mM for the same samples. The data were shifted vertically for clarity (Supplementary Fig. 26a shows the data without vertical shift). Error bars represent noise in the scattering measurements. b Correlation length, ξ, obtained from the Unified fit in the high-q region of the SAXS plots. The individual fits are shown in Supplementary Fig. 26b . Error bars obtained from fits were smaller than the markers and were not included. Their values can be found in Supplementary Table 11 .
Figure Legend Snippet: SAXS data of complex coacervate mimics samples. a Small-angle X-ray scattering plots of PDADMA/PAA complex coacervate samples prepared at 50–100 mM polymer charge and NaCl, with HEPES ranging between 25 and 50 mM, and MgCl 2 between 4.3 and 8.6 mM for the same samples. The data were shifted vertically for clarity (Supplementary Fig. 26a shows the data without vertical shift). Error bars represent noise in the scattering measurements. b Correlation length, ξ, obtained from the Unified fit in the high-q region of the SAXS plots. The individual fits are shown in Supplementary Fig. 26b . Error bars obtained from fits were smaller than the markers and were not included. Their values can be found in Supplementary Table 11 .

Techniques Used:

Changing the directionality of movements on the phase diagram. a Plot showing the change in salt concentrations in experiments that represent different compositions on the PDADMA/PAA phase diagram. HEPES and MgCl 2 were added at 2.5 and 0.43 mM initially and increased in all samples with the same factor of increase of NaCl on the y -axis. b Partitioning coefficient, taken as [U15] coacervate phase /[U15] dilute phase , with different concentrations of salt for the same polymer concentrations. c Whole-droplet FRAP experiments of the same range of samples. The inset corresponds to a zoom-in on the low concentration samples. Supplementary Table 12 includes all the fitting parameters. Partitioning data are averages and standard deviations obtained from three trials. Means and standard deviations in FRAP are obtained from three samples with two bleached droplets per sample. Individual data points are shown as grey markers.
Figure Legend Snippet: Changing the directionality of movements on the phase diagram. a Plot showing the change in salt concentrations in experiments that represent different compositions on the PDADMA/PAA phase diagram. HEPES and MgCl 2 were added at 2.5 and 0.43 mM initially and increased in all samples with the same factor of increase of NaCl on the y -axis. b Partitioning coefficient, taken as [U15] coacervate phase /[U15] dilute phase , with different concentrations of salt for the same polymer concentrations. c Whole-droplet FRAP experiments of the same range of samples. The inset corresponds to a zoom-in on the low concentration samples. Supplementary Table 12 includes all the fitting parameters. Partitioning data are averages and standard deviations obtained from three trials. Means and standard deviations in FRAP are obtained from three samples with two bleached droplets per sample. Individual data points are shown as grey markers.

Techniques Used: Concentration Assay

Drying and rehydrating controls the formation of PDADMA/PAA complex coacervates. a Scheme showing the motivation behind the wet-dry cycle experiment—an early Earth environmental scenario proposed for the emergence of chemical complexity. b Turbidity values for 1:1 PDADMA:PAA complex coacervates prepared at different charge and salt concentrations ([monomer charge] is calculated with respect to the concentration of the polymer repeat units). All samples contained 25 mM HEPES and 4.3 mM MgCl 2 . The plot on the left is a zoom-in on the low [charge] samples. Black markers represent turbidity below a cutoff of 20, which was chosen based on microscopy images (Supplementary Fig. 8 ). The width of all circles is proportional to the turbidity values of the overall samples (see legend above, T refers to transmittance). As a guide to the eye, a solid red line delimits the expected boundary between the two-phase and one-phase regions; the line is an approximate binodal curve. The charge concentration refers to one polymer type, not the total amount of both polymers. c Comparison between drying different 1:1 PDADMA:PAA coacervate compositions based on different starting locations on the phase diagram (i = filled triangle, ii = filled square, iii = filled diamond, with the open versions—open triangle, open square, open diamond—and dashed arrows showing the expected approximate 10× dehydration) in the time range of 0–160 min. Drying was performed in a heatblock at 95 °C; (i) shows the volume remaining at each time point (V), which is an average of 27 samples (see Fig. 2 ) and is applicable to (ii) and (iii) as they have the same drying rate. The dashed arrow in (ii) is expected to go up to 500 mM [NaCl] and 150 mM [polymer charge], and is thus not fully shown. (iv) Rehydration of (i) by adding a volume of water similar to the volume reached at each time point in reverse order of the top panel. All scale bars are 20 µm.
Figure Legend Snippet: Drying and rehydrating controls the formation of PDADMA/PAA complex coacervates. a Scheme showing the motivation behind the wet-dry cycle experiment—an early Earth environmental scenario proposed for the emergence of chemical complexity. b Turbidity values for 1:1 PDADMA:PAA complex coacervates prepared at different charge and salt concentrations ([monomer charge] is calculated with respect to the concentration of the polymer repeat units). All samples contained 25 mM HEPES and 4.3 mM MgCl 2 . The plot on the left is a zoom-in on the low [charge] samples. Black markers represent turbidity below a cutoff of 20, which was chosen based on microscopy images (Supplementary Fig. 8 ). The width of all circles is proportional to the turbidity values of the overall samples (see legend above, T refers to transmittance). As a guide to the eye, a solid red line delimits the expected boundary between the two-phase and one-phase regions; the line is an approximate binodal curve. The charge concentration refers to one polymer type, not the total amount of both polymers. c Comparison between drying different 1:1 PDADMA:PAA coacervate compositions based on different starting locations on the phase diagram (i = filled triangle, ii = filled square, iii = filled diamond, with the open versions—open triangle, open square, open diamond—and dashed arrows showing the expected approximate 10× dehydration) in the time range of 0–160 min. Drying was performed in a heatblock at 95 °C; (i) shows the volume remaining at each time point (V), which is an average of 27 samples (see Fig. 2 ) and is applicable to (ii) and (iii) as they have the same drying rate. The dashed arrow in (ii) is expected to go up to 500 mM [NaCl] and 150 mM [polymer charge], and is thus not fully shown. (iv) Rehydration of (i) by adding a volume of water similar to the volume reached at each time point in reverse order of the top panel. All scale bars are 20 µm.

Techniques Used: Concentration Assay, Microscopy

Partitioning of U15, a 15-mer oligouridylic acid, during PDADMA/PAA coacervate suspension drying. a Total volumes of PDADMA/PAA complex coacervate at each time point of drying. Averages and standard deviations are obtained from 9 independent trials ( N = 27). The starting composition is the same for all samples: [polymer charge] = [NaCl] = 5 mM, [HEPES] = 2.5 mM, [MgCl 2 ] = 0.43 mM. b Calculated increase in the concentrations of the solution components. Standard deviations are among the same trials used to calculate the volumes above. c To understand the partitioning of RNA within the coacervate while avoiding risk of hydrolysis during the temperature-induced drying, Alexa 647-tagged U15 was added after each drying step was complete and the solutions returned to room temperature. Fluorescent U15 was added such that its global concentration increased in the same way as all the other components in solution had, during the drying steps. Overall concentrations of U15 were as low as 0.02 µM and never increased above 0.2 µM, which is around 25,000x less than the lowest polymer concentrations, and is not believed to interfere with phase separation. d The partitioning coefficient, calculated as [U15] coacervate phase /[U15] dilute phase , and U15 concentrations within the coacervate after addition of the fluorescent RNA in equal numbers of moles. Means and standard deviations are obtained from three trials with analysis of 15 droplets over five images, per trial. e Confocal fluorescence microscopy images showing the partitioned U15 within the droplets. While the partitioning coefficients decrease, the internal concentrations do not change significantly as quantification in d and Supplementary Fig. 14 show. Individual data points are shown in Supplementary Fig. 14 .
Figure Legend Snippet: Partitioning of U15, a 15-mer oligouridylic acid, during PDADMA/PAA coacervate suspension drying. a Total volumes of PDADMA/PAA complex coacervate at each time point of drying. Averages and standard deviations are obtained from 9 independent trials ( N = 27). The starting composition is the same for all samples: [polymer charge] = [NaCl] = 5 mM, [HEPES] = 2.5 mM, [MgCl 2 ] = 0.43 mM. b Calculated increase in the concentrations of the solution components. Standard deviations are among the same trials used to calculate the volumes above. c To understand the partitioning of RNA within the coacervate while avoiding risk of hydrolysis during the temperature-induced drying, Alexa 647-tagged U15 was added after each drying step was complete and the solutions returned to room temperature. Fluorescent U15 was added such that its global concentration increased in the same way as all the other components in solution had, during the drying steps. Overall concentrations of U15 were as low as 0.02 µM and never increased above 0.2 µM, which is around 25,000x less than the lowest polymer concentrations, and is not believed to interfere with phase separation. d The partitioning coefficient, calculated as [U15] coacervate phase /[U15] dilute phase , and U15 concentrations within the coacervate after addition of the fluorescent RNA in equal numbers of moles. Means and standard deviations are obtained from three trials with analysis of 15 droplets over five images, per trial. e Confocal fluorescence microscopy images showing the partitioned U15 within the droplets. While the partitioning coefficients decrease, the internal concentrations do not change significantly as quantification in d and Supplementary Fig. 14 show. Individual data points are shown in Supplementary Fig. 14 .

Techniques Used: Concentration Assay, Fluorescence, Microscopy

Impact of repeating wet-dry cycles on the complex coacervate. a PDADMA/PAA coacervates ([Charge] = [NaCl] = 10 mM, [HEPES] = 5 mM, [MgCl 2 ] = 0.9 mM) were fully dried and rehydrated for five consecutive cycles. The plot shows the average weights after drying-rehydration. b The concentration of U15 in the same coacervate after rehydration of samples obtained by confocal microscopy using Alexa 647-U15 as a probe. c The partitioning coefficient, K, taken as [U15] coacervate phase /[U15] dilute phase , after each cycle. d Apparent diffusion coefficients obtained from halftime of fluorescence recovery during FRAP experiment where whole coacervate droplets, containing the Alexa 647-U15 probe, were bleached. e Apparent diffusion coefficients in FRAP experiments where a 1-µm diameter region within the coacervate phase was bleached. The data in b – e are averages and standard deviations of two trials with an analysis of 10 droplets total for b , with the same values used when dividing by the [U15] in the dilute phase in c , six droplets in d , and 10 intradroplet areas in e . Individual data points are shown as gray markers. Supplementary Tables 5 and 6 include all FRAP fitting parameters.
Figure Legend Snippet: Impact of repeating wet-dry cycles on the complex coacervate. a PDADMA/PAA coacervates ([Charge] = [NaCl] = 10 mM, [HEPES] = 5 mM, [MgCl 2 ] = 0.9 mM) were fully dried and rehydrated for five consecutive cycles. The plot shows the average weights after drying-rehydration. b The concentration of U15 in the same coacervate after rehydration of samples obtained by confocal microscopy using Alexa 647-U15 as a probe. c The partitioning coefficient, K, taken as [U15] coacervate phase /[U15] dilute phase , after each cycle. d Apparent diffusion coefficients obtained from halftime of fluorescence recovery during FRAP experiment where whole coacervate droplets, containing the Alexa 647-U15 probe, were bleached. e Apparent diffusion coefficients in FRAP experiments where a 1-µm diameter region within the coacervate phase was bleached. The data in b – e are averages and standard deviations of two trials with an analysis of 10 droplets total for b , with the same values used when dividing by the [U15] in the dilute phase in c , six droplets in d , and 10 intradroplet areas in e . Individual data points are shown as gray markers. Supplementary Tables 5 and 6 include all FRAP fitting parameters.

Techniques Used: Concentration Assay, Confocal Microscopy, Diffusion-based Assay, Fluorescence

Dynamics of U15 in the complex coacervate during drying. a Fluorescence recovery after photobleaching, FRAP, over a whole 1:1 PDADMA:PAA coacervate droplet in solutions dried at 95 °C for 160 min and allowed to return to room temperature before partitioning Alexa 647-U15 within. b FRAP over part of the same coacervate in solutions dried at the same temperature for 80 min and allowed to return to room temperature before adding U15. c Recovery of the fluorescence after whole-droplet photobleaching of U15 partitioned within a coacervate in mixtures allowed to dry for 160 min. d Recovery of the fluorescence after partial-droplet photobleaching of U15 partitioned within a coacervate in mixtures allowed to dry for 80 min. Circle markers represent corrected fluorescence intensity in c, d . Lines are fitting results according to equations mentioned in the Methods section. e Apparent diffusion coefficients and recovery halftimes of U15 as it recovers after bleaching of whole coacervate droplets at each time point of the drying experiment. f Apparent diffusion coefficient and recovery halftimes of U15 as it recovers after bleaching of partial droplets at each time point. The bars are averages of five trials with at least two droplets each in e . In f , the bars are averages of three trials with at least three droplets analyzed in each. All individual data points are shown in Supplementary Tables 2 and 3 , and as gray markers for the apparent diffusion coefficients in e and f . The error bars represent the standard deviations among all trials.
Figure Legend Snippet: Dynamics of U15 in the complex coacervate during drying. a Fluorescence recovery after photobleaching, FRAP, over a whole 1:1 PDADMA:PAA coacervate droplet in solutions dried at 95 °C for 160 min and allowed to return to room temperature before partitioning Alexa 647-U15 within. b FRAP over part of the same coacervate in solutions dried at the same temperature for 80 min and allowed to return to room temperature before adding U15. c Recovery of the fluorescence after whole-droplet photobleaching of U15 partitioned within a coacervate in mixtures allowed to dry for 160 min. d Recovery of the fluorescence after partial-droplet photobleaching of U15 partitioned within a coacervate in mixtures allowed to dry for 80 min. Circle markers represent corrected fluorescence intensity in c, d . Lines are fitting results according to equations mentioned in the Methods section. e Apparent diffusion coefficients and recovery halftimes of U15 as it recovers after bleaching of whole coacervate droplets at each time point of the drying experiment. f Apparent diffusion coefficient and recovery halftimes of U15 as it recovers after bleaching of partial droplets at each time point. The bars are averages of five trials with at least two droplets each in e . In f , the bars are averages of three trials with at least three droplets analyzed in each. All individual data points are shown in Supplementary Tables 2 and 3 , and as gray markers for the apparent diffusion coefficients in e and f . The error bars represent the standard deviations among all trials.

Techniques Used: Fluorescence, Diffusion-based Assay

12) Product Images from "Recruitment of Cellular Recombination and Repair Proteins to Sites of Herpes Simplex Virus Type 1 DNA Replication Is Dependent on the Composition of Viral Proteins within Prereplicative Sites and Correlates with the Induction of the DNA Damage Response"

Article Title: Recruitment of Cellular Recombination and Repair Proteins to Sites of Herpes Simplex Virus Type 1 DNA Replication Is Dependent on the Composition of Viral Proteins within Prereplicative Sites and Correlates with the Induction of the DNA Damage Response

Journal: Journal of Virology

doi: 10.1128/JVI.78.9.4783-4796.2004

Western blots analysis. Lysates were prepared at the indicated hours postinfection (h.p.i.). In the presence or absence of PAA, Vero cells were either mock-infected or infected with either KOS or the polymerase null mutant virus HP66. As a control for DNA damage, mock-infected cells were also treated with 1 μM camptothecin (CPT) for 6 h prior to harvest. (A) Membranes were probed with antibodies directed against UL29, NBS1, Ku86, RAD51, and RPA32 as described in Materials and Methods. The open arrows indicate the slower-migrating species of NBS1 and RPA32. Results for tubulin are provided as a loading control. (B) Membranes were probed with antibodies directed against RAD52, RAD50, MRE11, and DNA-PKcs.
Figure Legend Snippet: Western blots analysis. Lysates were prepared at the indicated hours postinfection (h.p.i.). In the presence or absence of PAA, Vero cells were either mock-infected or infected with either KOS or the polymerase null mutant virus HP66. As a control for DNA damage, mock-infected cells were also treated with 1 μM camptothecin (CPT) for 6 h prior to harvest. (A) Membranes were probed with antibodies directed against UL29, NBS1, Ku86, RAD51, and RPA32 as described in Materials and Methods. The open arrows indicate the slower-migrating species of NBS1 and RPA32. Results for tubulin are provided as a loading control. (B) Membranes were probed with antibodies directed against RAD52, RAD50, MRE11, and DNA-PKcs.

Techniques Used: Western Blot, Infection, Mutagenesis, Cycling Probe Technology

13) Product Images from "Herpes Simplex Virus 2 Infection Impacts Stress Granule Accumulation"

Article Title: Herpes Simplex Virus 2 Infection Impacts Stress Granule Accumulation

Journal: Journal of Virology

doi: 10.1128/JVI.00313-12

SG accumulation in HSV-2-infected cells in the presence of PAA. (A) HeLa cells were infected with HSV-2 strain HG52 at an MOI of 5 in the presence or absence of 200 μg of phosphonoacetic acid/ml (+PAA or −PAA, respectively). At the indicated
Figure Legend Snippet: SG accumulation in HSV-2-infected cells in the presence of PAA. (A) HeLa cells were infected with HSV-2 strain HG52 at an MOI of 5 in the presence or absence of 200 μg of phosphonoacetic acid/ml (+PAA or −PAA, respectively). At the indicated

Techniques Used: Infection

14) Product Images from "Graphene Wrapping of Electrospun Nanofibers for Enhanced Electrochemical Sensing"

Article Title: Graphene Wrapping of Electrospun Nanofibers for Enhanced Electrochemical Sensing

Journal: ACS Omega

doi: 10.1021/acsomega.0c05823

Raman spectrum of rGO-wrapped PAN/PAA electrospun fibers.
Figure Legend Snippet: Raman spectrum of rGO-wrapped PAN/PAA electrospun fibers.

Techniques Used:

Annotated FTIR spectrum of the graphene-wrapped electrospun nanofibers. The presence of C–H, C≡N, C=O, C=C, C–OH, C–C, and C–O bonds can be identified. The C=C and C–O bonds (red) are attributable to the graphene wrap, while the rest (blue) are attributable to the PAN and PAA polymers of the nanofibers.
Figure Legend Snippet: Annotated FTIR spectrum of the graphene-wrapped electrospun nanofibers. The presence of C–H, C≡N, C=O, C=C, C–OH, C–C, and C–O bonds can be identified. The C=C and C–O bonds (red) are attributable to the graphene wrap, while the rest (blue) are attributable to the PAN and PAA polymers of the nanofibers.

Techniques Used:

SEM images of (a) typical electrospun PAN/PAA nanofibers and (b–f) the graphene wrapping process. (b) Faint layers of rGO encapsulating fibers. (c) An example of a graphene-wrapped “yarn” of fibers, which is most likely a consequence of the constant stirring required in the graphene wrapping method. (d) The graphene wrap is agglomerated as sheets instead of encapsulating individual fibers in areas of a high fiber density. (e) Smooth nanofiber morphology after carbonization. (f) Surface texture of the graphene-wrapped carbonized nanofibers.
Figure Legend Snippet: SEM images of (a) typical electrospun PAN/PAA nanofibers and (b–f) the graphene wrapping process. (b) Faint layers of rGO encapsulating fibers. (c) An example of a graphene-wrapped “yarn” of fibers, which is most likely a consequence of the constant stirring required in the graphene wrapping method. (d) The graphene wrap is agglomerated as sheets instead of encapsulating individual fibers in areas of a high fiber density. (e) Smooth nanofiber morphology after carbonization. (f) Surface texture of the graphene-wrapped carbonized nanofibers.

Techniques Used:

15) Product Images from "Influence of gold nanoparticle surface chemistry and diameter upon Alzheimer’s disease amyloid-β protein aggregation"

Article Title: Influence of gold nanoparticle surface chemistry and diameter upon Alzheimer’s disease amyloid-β protein aggregation

Journal: Journal of Biological Engineering

doi: 10.1186/s13036-017-0047-6

Toxicity of surface-coated gold NPs. SH-SY5Y human neuroblastoma cells were incubated (24 h) alone ( negative control ) or in the presence of 100 pM ( closed bars ) or 200 pM ( open bars ) NPs. a Treatment with 18 nm NPs coated with citrate, CTAB, PAA, or PAH. b Treatment with 8 nm, 18 nm, or 40 nm NPs coated with PAA. Treatment with 2% Triton-X (Tr-X) served as a positive control. Cellular viability was assessed using XTT reduction. Results are shown as the percentage of viable cells relative to the negative control and represent the mean of 2–3 independent experiments, performed with 6 replicates. Error bars represent SEM. * p
Figure Legend Snippet: Toxicity of surface-coated gold NPs. SH-SY5Y human neuroblastoma cells were incubated (24 h) alone ( negative control ) or in the presence of 100 pM ( closed bars ) or 200 pM ( open bars ) NPs. a Treatment with 18 nm NPs coated with citrate, CTAB, PAA, or PAH. b Treatment with 8 nm, 18 nm, or 40 nm NPs coated with PAA. Treatment with 2% Triton-X (Tr-X) served as a positive control. Cellular viability was assessed using XTT reduction. Results are shown as the percentage of viable cells relative to the negative control and represent the mean of 2–3 independent experiments, performed with 6 replicates. Error bars represent SEM. * p

Techniques Used: Incubation, Negative Control, Positive Control

Effect of surface-coated gold NPs on ThT fluorescence detection of Aβ 1-40 aggregates. Aβ 1–40 fibrils diluted in 40 mM Tris–HCl (pH 8.0) to a final concentration of 5 μM were combined with 8.75 μM ThT and incubated (2 h) alone (control) or with 5 pM, 10 pM, 20 pM, 50 pM, 100 pM, or 200 pM NPs. a Incubation with 18 nm NPs displaying citrate (■), CTAB (●), PAA (▲), or PAH (♦) surface chemistries. b Incubation with 8 nm (▶), 18 nm (▲), or 40 nm (◀) NPs coated with PAA. ThT fluorescence was evaluated, and results are expressed as the fraction of ThT fluorescence observed for the control. Error bars represent SEM, n = 3. † p
Figure Legend Snippet: Effect of surface-coated gold NPs on ThT fluorescence detection of Aβ 1-40 aggregates. Aβ 1–40 fibrils diluted in 40 mM Tris–HCl (pH 8.0) to a final concentration of 5 μM were combined with 8.75 μM ThT and incubated (2 h) alone (control) or with 5 pM, 10 pM, 20 pM, 50 pM, 100 pM, or 200 pM NPs. a Incubation with 18 nm NPs displaying citrate (■), CTAB (●), PAA (▲), or PAH (♦) surface chemistries. b Incubation with 8 nm (▶), 18 nm (▲), or 40 nm (◀) NPs coated with PAA. ThT fluorescence was evaluated, and results are expressed as the fraction of ThT fluorescence observed for the control. Error bars represent SEM, n = 3. † p

Techniques Used: Fluorescence, Concentration Assay, Incubation

Effect of surface-coated gold NPs with different surface coatings and diameters on Aβ 1-40 aggregate morphology. Aβ 1–40 monomer diluted to 40 μM in 40 mM Tris–HCl (pH 8.0) was aggregated alone (control, panels a , d ) or in the presence of 200 pM citrate-coated (panel b ), PAH-coated (panel c ), CTAB-coated (panel e ), or PAA-coated (panel g ) NPs 18 nm in diameter. Additionally, monomer was aggregated in the presence of 200 pM PAA-coated NPs exhibiting diameters of 8 nm (panel f ) or 40 nm (panel h ). Monomer aggregation was induced by continuous agitation, and the control reaction was monitored periodically via ThT fluorescence. Upon evidence of equilibrium, samples were gridded and visualized by TEM. Results are representative of 2 independent experiments. Images are shown relative to a scale bar of 500 nm at 100,000x (panels a - c , e - g ) or 75,000x (panels d , h ) magnification
Figure Legend Snippet: Effect of surface-coated gold NPs with different surface coatings and diameters on Aβ 1-40 aggregate morphology. Aβ 1–40 monomer diluted to 40 μM in 40 mM Tris–HCl (pH 8.0) was aggregated alone (control, panels a , d ) or in the presence of 200 pM citrate-coated (panel b ), PAH-coated (panel c ), CTAB-coated (panel e ), or PAA-coated (panel g ) NPs 18 nm in diameter. Additionally, monomer was aggregated in the presence of 200 pM PAA-coated NPs exhibiting diameters of 8 nm (panel f ) or 40 nm (panel h ). Monomer aggregation was induced by continuous agitation, and the control reaction was monitored periodically via ThT fluorescence. Upon evidence of equilibrium, samples were gridded and visualized by TEM. Results are representative of 2 independent experiments. Images are shown relative to a scale bar of 500 nm at 100,000x (panels a - c , e - g ) or 75,000x (panels d , h ) magnification

Techniques Used: Fluorescence, Transmission Electron Microscopy

Effect of 18 nm surface-coated gold NPs on Aβ 1–40 monomer aggregation. Aβ 1–40 monomer diluted to 40 μM in 40 mM Tris–HCl (pH 8.0) was aggregated alone (control, Ο) or in the presence of surface-coated NPs. a Aggregation in the presence of 200 pM NPs displaying citrate (■), PAA (▲), or PAH (♦) surface chemistries. b Aggregation in the presence of 200 pM (♦), 100 pM ( ), or 20 pM ( ) PAH-coated NPs. Monomer aggregation was induced by continuous agitation and monitored periodically via ThT fluorescence. ThT fluorescence, expressed as the fraction of the control equilibrium plateau, is plotted versus relative time, which is the fraction of the control lag time. Results are representative of 3–5 independent experiments
Figure Legend Snippet: Effect of 18 nm surface-coated gold NPs on Aβ 1–40 monomer aggregation. Aβ 1–40 monomer diluted to 40 μM in 40 mM Tris–HCl (pH 8.0) was aggregated alone (control, Ο) or in the presence of surface-coated NPs. a Aggregation in the presence of 200 pM NPs displaying citrate (■), PAA (▲), or PAH (♦) surface chemistries. b Aggregation in the presence of 200 pM (♦), 100 pM ( ), or 20 pM ( ) PAH-coated NPs. Monomer aggregation was induced by continuous agitation and monitored periodically via ThT fluorescence. ThT fluorescence, expressed as the fraction of the control equilibrium plateau, is plotted versus relative time, which is the fraction of the control lag time. Results are representative of 3–5 independent experiments

Techniques Used: Fluorescence

16) Product Images from "A potent novel vaccine adjuvant based on straight polyacrylate"

Article Title: A potent novel vaccine adjuvant based on straight polyacrylate

Journal: International Journal of Pharmaceutics: X

doi: 10.1016/j.ijpx.2020.100054

Adjuvant activity of different technical lots of PAA is dependent on polymer size and dose and not on polymer branching. C57BL/6J mice (n = 10/group) were immunized 2 times, 25 days apart, by IM injection of 2 μg of recombinant CMV-gB without adjuvant or with MF59 or different PAAs at increasing concentrations (25-50-100-200-400 μg/dose) under a final volume of 50 μl. The PAAs used in this experiment were lots # 566253 (○), #652979 (□), #240413 (∆) and #655555 (◊) characterized in Table 1 . Two weeks following the second administration (D39), CMV neutralizing titers determined on ARPE-19 cells (Panel A), CMV-gB-specific IgG1 (panel B) and IgG2c (panel C) titers and spleen cell IFN-γ responses (panel D) were determined as in Fig. 1 . Antibody and IFN-γ levels obtained in mice injected with CMV-gB alone or combined with MF59, are shown as dotted and dash lines, respectively. No IL-5 was detected in the PAA groups in this study.
Figure Legend Snippet: Adjuvant activity of different technical lots of PAA is dependent on polymer size and dose and not on polymer branching. C57BL/6J mice (n = 10/group) were immunized 2 times, 25 days apart, by IM injection of 2 μg of recombinant CMV-gB without adjuvant or with MF59 or different PAAs at increasing concentrations (25-50-100-200-400 μg/dose) under a final volume of 50 μl. The PAAs used in this experiment were lots # 566253 (○), #652979 (□), #240413 (∆) and #655555 (◊) characterized in Table 1 . Two weeks following the second administration (D39), CMV neutralizing titers determined on ARPE-19 cells (Panel A), CMV-gB-specific IgG1 (panel B) and IgG2c (panel C) titers and spleen cell IFN-γ responses (panel D) were determined as in Fig. 1 . Antibody and IFN-γ levels obtained in mice injected with CMV-gB alone or combined with MF59, are shown as dotted and dash lines, respectively. No IL-5 was detected in the PAA groups in this study.

Techniques Used: Activity Assay, Mouse Assay, Injection, Recombinant

Adjuvant activity of PAA is strongly dependent on polymer size. C57BL/6J mice ( n = 10/group) were immunized 2 times, 4 weeks apart, by IM injection of 2 μg of recombinant CMV-gB without adjuvant (white bars) or with MF59 (red bars) or PAA (blue bars) under a final volume of 50 μl. Low Mw PAA (lot#617193; Mw = 9 kDa and PI = 3.1), medium Mw PAA (lot#601318; Mw = 134 kDa and PI = 2.4) and high molecular weight PAA (lot#566253; Mw = 489 kDa and PI = 3.8) were all tested at 200 μg/dose. Two weeks after the second immunization (D42), specific serum antibody responses and spleen cell cytokine responses were determined as described in the Materials and Methods section. Panel A: CMV neutralizing antibody titers measured on ARPE-19 epithelial cells. Panel B: CMV neutralizing antibody titers measured on MRC-5 fibroblasts. Panel C: CMV-gB-specific IgG1 and IgG2c titers. Panel D: IL-5 and IFN-γ productions from restimulated splenocytes cultures. Titers were plotted as geometric mean titers with 95% Confidence Interval (CI) on a log scale. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
Figure Legend Snippet: Adjuvant activity of PAA is strongly dependent on polymer size. C57BL/6J mice ( n = 10/group) were immunized 2 times, 4 weeks apart, by IM injection of 2 μg of recombinant CMV-gB without adjuvant (white bars) or with MF59 (red bars) or PAA (blue bars) under a final volume of 50 μl. Low Mw PAA (lot#617193; Mw = 9 kDa and PI = 3.1), medium Mw PAA (lot#601318; Mw = 134 kDa and PI = 2.4) and high molecular weight PAA (lot#566253; Mw = 489 kDa and PI = 3.8) were all tested at 200 μg/dose. Two weeks after the second immunization (D42), specific serum antibody responses and spleen cell cytokine responses were determined as described in the Materials and Methods section. Panel A: CMV neutralizing antibody titers measured on ARPE-19 epithelial cells. Panel B: CMV neutralizing antibody titers measured on MRC-5 fibroblasts. Panel C: CMV-gB-specific IgG1 and IgG2c titers. Panel D: IL-5 and IFN-γ productions from restimulated splenocytes cultures. Titers were plotted as geometric mean titers with 95% Confidence Interval (CI) on a log scale. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Techniques Used: Activity Assay, Mouse Assay, Injection, Recombinant, Molecular Weight

17) Product Images from "The Human Cytomegalovirus Gene Products Essential for Late Viral Gene Expression Assemble into Prereplication Complexes before Viral DNA Replication ▿"

Article Title: The Human Cytomegalovirus Gene Products Essential for Late Viral Gene Expression Assemble into Prereplication Complexes before Viral DNA Replication ▿

Journal: Journal of Virology

doi: 10.1128/JVI.00384-11

UL95-HA, UL87-myc, and Flag-UL79 fusion proteins are expressed at early times after infection. Viral DNA synthesis was inhibited with PAA or GCV. Western blot analysis was performed using an antibody against immediate-early proteins pIE86 and pIE72 (UL122
Figure Legend Snippet: UL95-HA, UL87-myc, and Flag-UL79 fusion proteins are expressed at early times after infection. Viral DNA synthesis was inhibited with PAA or GCV. Western blot analysis was performed using an antibody against immediate-early proteins pIE86 and pIE72 (UL122

Techniques Used: Infection, DNA Synthesis, Western Blot

18) Product Images from "13C NMR isotopomer analysis reveals a connection between pyruvate cycling and glucose-stimulated insulin secretion (GSIS)"

Article Title: 13C NMR isotopomer analysis reveals a connection between pyruvate cycling and glucose-stimulated insulin secretion (GSIS)

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.052005699

A stimulatory substrate (DMM) and an inhibitor (PAA) of pyruvate cycling have proportional effects on insulin secretion. Insulin secretion was measured from 832/13 cells during incubation with 12 mM [U- 13 C 6 ]glucose. A shows insulin secretion and pyruvate cycling in the presence and absence of 10 mM DMM. B shows insulin secretion and pyruvate cycling in the presence and absence of 5 mM PAA, an inhibitor of PC. Data represent the mean ± SE for 4 independent determinations.
Figure Legend Snippet: A stimulatory substrate (DMM) and an inhibitor (PAA) of pyruvate cycling have proportional effects on insulin secretion. Insulin secretion was measured from 832/13 cells during incubation with 12 mM [U- 13 C 6 ]glucose. A shows insulin secretion and pyruvate cycling in the presence and absence of 10 mM DMM. B shows insulin secretion and pyruvate cycling in the presence and absence of 5 mM PAA, an inhibitor of PC. Data represent the mean ± SE for 4 independent determinations.

Techniques Used: Incubation

; in order of increasing capacity for GSIS). The open diamond represents 832/13 cells incubated with 12 mM glucose plus 10 mM DMM, a stimulatory metabolite for pyruvate cycling. The open triangle represents 832/13 cells incubated with 12 mM glucose plus 5 mM PAA, an inhibitor of PC and pyruvate cycling.
Figure Legend Snippet: ; in order of increasing capacity for GSIS). The open diamond represents 832/13 cells incubated with 12 mM glucose plus 10 mM DMM, a stimulatory metabolite for pyruvate cycling. The open triangle represents 832/13 cells incubated with 12 mM glucose plus 5 mM PAA, an inhibitor of PC and pyruvate cycling.

Techniques Used: Incubation

19) Product Images from "Adherence of the Gram-Positive Bacterium Ruminococcus albus to Cellulose and Identification of a Novel Form of Cellulose-Binding Protein Which Belongs to the Pil Family of Proteins †"

Article Title: Adherence of the Gram-Positive Bacterium Ruminococcus albus to Cellulose and Identification of a Novel Form of Cellulose-Binding Protein Which Belongs to the Pil Family of Proteins †

Journal: Journal of Bacteriology

doi:

Northern blot analysis of cbpC transcript abundance following growth in various media. Total RNA was harvested from cells following growth in cellobiose medium containing either ruminal fluid (lane 1), no additions (lane 2), or both PAA and PPA. (A) Results obtained with the cbpC -gene specific probe; (B) the same membrane probed with an oligonucleotide complementary to 16S rRNA.
Figure Legend Snippet: Northern blot analysis of cbpC transcript abundance following growth in various media. Total RNA was harvested from cells following growth in cellobiose medium containing either ruminal fluid (lane 1), no additions (lane 2), or both PAA and PPA. (A) Results obtained with the cbpC -gene specific probe; (B) the same membrane probed with an oligonucleotide complementary to 16S rRNA.

Techniques Used: Northern Blot

20) Product Images from "The Efficacy of Sodium Bisulfate Salt (SBS) Alone and Combined With Peracetic Acid (PAA) as an Antimicrobial on Whole Chicken Drumsticks Artificially Inoculated With Salmonella Enteritidis"

Article Title: The Efficacy of Sodium Bisulfate Salt (SBS) Alone and Combined With Peracetic Acid (PAA) as an Antimicrobial on Whole Chicken Drumsticks Artificially Inoculated With Salmonella Enteritidis

Journal: Frontiers in Veterinary Science

doi: 10.3389/fvets.2019.00006

The linear effect of Sodium Bisulfate, SBS, as an antimicrobial 15 s part dip on suppressing the population of Salmonella Enteritidis on whole chicken drumsticks on d 0, 1, and 3. Drumsticks were artificially inoculated with 10 7 CFU/g of S . Enteritidis and subsequently treated in 300 mL of antimicrobial treatments to identify the remaining population of Salmonella . There were eight antimicrobial treatments consisting of: a no treatment Control, tap water (TW), tap water with the addition of either 1, 2, or 3% SBS indicated as TW+SBS1, TW+SBS2, and TW+SBS3, and the combination of 1, 2, and 3% SBS with 200 ppm of peracetic acid (PAA), represented as TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA. In the current figure, a linear trend was investigated for the incremental increase in SBS concentration, Control ( n = 4), TW ( n = 4), TW+SBS1 (TW+SBS1 and TW+SBS1+PAA, n = 8), TW+SBS2 (TW+SBS2 and TW+SBS2+PAA, n = 8), and TW+SBS3 (TW+SBS3 and TW+SBS3+PAA, n = 8), over a 3-d refrigeration period at 4°C. Individual SEM for Control, TW, TW+SBS1, TW+SBS2, and TW+SBS3 was 0.292, 0.284, 0.281, 0.229, 0.146 log CFU/g for d 0, 0.205, 0.309, 0.092, 0.122, and 0.089 log CFU/g for d 1, and 0.223, 0.278, 0.082, 0.131, and 0.067 for d 3, respectively. Pooled SEM for d 0 is 0.227, 0.196 for d 1, and 0.175 for d 3; N = 32. Means with different superscripts are considered different (a–d).
Figure Legend Snippet: The linear effect of Sodium Bisulfate, SBS, as an antimicrobial 15 s part dip on suppressing the population of Salmonella Enteritidis on whole chicken drumsticks on d 0, 1, and 3. Drumsticks were artificially inoculated with 10 7 CFU/g of S . Enteritidis and subsequently treated in 300 mL of antimicrobial treatments to identify the remaining population of Salmonella . There were eight antimicrobial treatments consisting of: a no treatment Control, tap water (TW), tap water with the addition of either 1, 2, or 3% SBS indicated as TW+SBS1, TW+SBS2, and TW+SBS3, and the combination of 1, 2, and 3% SBS with 200 ppm of peracetic acid (PAA), represented as TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA. In the current figure, a linear trend was investigated for the incremental increase in SBS concentration, Control ( n = 4), TW ( n = 4), TW+SBS1 (TW+SBS1 and TW+SBS1+PAA, n = 8), TW+SBS2 (TW+SBS2 and TW+SBS2+PAA, n = 8), and TW+SBS3 (TW+SBS3 and TW+SBS3+PAA, n = 8), over a 3-d refrigeration period at 4°C. Individual SEM for Control, TW, TW+SBS1, TW+SBS2, and TW+SBS3 was 0.292, 0.284, 0.281, 0.229, 0.146 log CFU/g for d 0, 0.205, 0.309, 0.092, 0.122, and 0.089 log CFU/g for d 1, and 0.223, 0.278, 0.082, 0.131, and 0.067 for d 3, respectively. Pooled SEM for d 0 is 0.227, 0.196 for d 1, and 0.175 for d 3; N = 32. Means with different superscripts are considered different (a–d).

Techniques Used: Concentration Assay

The comparative effect of 3% Sodium Bisulfate, SBS, and 200 ppm of peracetic acid, PAA, utilized alone or in combination as antimicrobial 15 s part dips on the presence of Salmonella Enteritidis on whole chicken drumsticks on d 0, 1, and 3. In the current study, drumsticks were artificially inoculated with 10 7 CFU/g of S . Enteritidis and subsequently treated in 300 mL of antimicrobial treatments to identify the remaining population of Salmonella . In the study, there were eight treatments, consisting of: a no treatment Control, tap water (TW), tap water with the addition of either 1, 2, or 3% sodium bisulfate (SBS) indicated as TW+SBS1, TW+SBS2, and TW+SBS3, and the combination of 1, 2, and 3% SBS with 200 ppm of peracetic acid (PAA), represented as TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA. However, in the current figure only the Control, TW, TW+SBS3, and TW+SBS3+PAA is represented and is separated by d 0, 1, and 3 of 4°C incubation. Individual SEM for Control, TW, TW+SBS3, and TW+SBS3+PAA was 0.293, 0.284, 0.205, and 0.232 for d 0; 0.205, 0.309, 0.112, and 0.143 for d 1; and 0.223, 0.278, 0.119, and 0.083 for d 3, respectively. F -test P -values are 0.0169 for d 0, 0.0001 for d 1, and
Figure Legend Snippet: The comparative effect of 3% Sodium Bisulfate, SBS, and 200 ppm of peracetic acid, PAA, utilized alone or in combination as antimicrobial 15 s part dips on the presence of Salmonella Enteritidis on whole chicken drumsticks on d 0, 1, and 3. In the current study, drumsticks were artificially inoculated with 10 7 CFU/g of S . Enteritidis and subsequently treated in 300 mL of antimicrobial treatments to identify the remaining population of Salmonella . In the study, there were eight treatments, consisting of: a no treatment Control, tap water (TW), tap water with the addition of either 1, 2, or 3% sodium bisulfate (SBS) indicated as TW+SBS1, TW+SBS2, and TW+SBS3, and the combination of 1, 2, and 3% SBS with 200 ppm of peracetic acid (PAA), represented as TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA. However, in the current figure only the Control, TW, TW+SBS3, and TW+SBS3+PAA is represented and is separated by d 0, 1, and 3 of 4°C incubation. Individual SEM for Control, TW, TW+SBS3, and TW+SBS3+PAA was 0.293, 0.284, 0.205, and 0.232 for d 0; 0.205, 0.309, 0.112, and 0.143 for d 1; and 0.223, 0.278, 0.119, and 0.083 for d 3, respectively. F -test P -values are 0.0169 for d 0, 0.0001 for d 1, and

Techniques Used: Incubation

The effect of Sodium Bisulfate, SBS, and 200 ppm of peracetic acid, PAA, utilized alone or in combination as an antimicrobial 15 s part dip on the population of Salmonella Enteritidis on whole chicken drumsticks. In the current study, drumsticks were artificially inoculated with 10 7 CFU/g of S . Enteritidis and subsequently treated in 300 mL of antimicrobial treatments to identify the remaining population of Salmonella . There were eight treatments consisting of: a no treatment Control, tap water (TW), tap water with the addition of either 1, 2, or 3% SBS indicated as TW+SBS1, TW+SBS2, and TW+SBS3, and the combination of 1, 2, and 3% SBS with 200 ppm of peracetic acid (PAA), represented as TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA. The current figure demonstrates the effect the treatments had on Salmonella population regardless of refrigeration (4°C) time, d 0, 1, and 3. Individual standard error of the mean (SEM) for Control, TW, TW+SBS1, TW+SBS2, TW+SBS3, TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA were 0.136, 0.180, 0.089, 0.066, 0.091, 0.190, 0.132, 0.087 log CFU/g, respectively. F -test P -value
Figure Legend Snippet: The effect of Sodium Bisulfate, SBS, and 200 ppm of peracetic acid, PAA, utilized alone or in combination as an antimicrobial 15 s part dip on the population of Salmonella Enteritidis on whole chicken drumsticks. In the current study, drumsticks were artificially inoculated with 10 7 CFU/g of S . Enteritidis and subsequently treated in 300 mL of antimicrobial treatments to identify the remaining population of Salmonella . There were eight treatments consisting of: a no treatment Control, tap water (TW), tap water with the addition of either 1, 2, or 3% SBS indicated as TW+SBS1, TW+SBS2, and TW+SBS3, and the combination of 1, 2, and 3% SBS with 200 ppm of peracetic acid (PAA), represented as TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA. The current figure demonstrates the effect the treatments had on Salmonella population regardless of refrigeration (4°C) time, d 0, 1, and 3. Individual standard error of the mean (SEM) for Control, TW, TW+SBS1, TW+SBS2, TW+SBS3, TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA were 0.136, 0.180, 0.089, 0.066, 0.091, 0.190, 0.132, 0.087 log CFU/g, respectively. F -test P -value

Techniques Used:

The comparative effect of 1% Sodium Bisulfate, SBS, and 200 ppm of peracetic acid, PAA, utilized alone or in combination as antimicrobial 15 s part dips on the presence of Salmonella Enteritidis on whole chicken drumsticks on d 0, 1, and 3. In the current study, drumsticks were artificially inoculated with 10 7 CFU/g of S . Enteritidis and subsequently treated in 300 mL of antimicrobial treatments to identify the remaining population of Salmonella . In the study, there were eight treatments, consisting of: a no treatment Control, tap water (TW), tap water with the addition of either 1, 2, or 3% SBS indicated as TW+SBS1, TW+SBS2, and TW+SBS3, and the combination of 1, 2, and 3% SBS with 200 ppm of peracetic acid (PAA), represented as TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA. However, in the current figure only the Control, TW, TW+SBS1, and TW+SBS1+PAA is represented and is separated by d 0, 1, and 3 of 4°C incubation. Individual SEM for Control, TW, TW+SBS1, and TW+SBS1+PAA was 0.293, 0.284, 0.196, and 0.432 for d 0; 0.205, 0.309, 0.150, and 0.086 for d 1; and 0.223, 0.278, 0.064, and 0.078 for d 3, respectively. P -value for d 0 is 0.0116, 0.0067 for d 1, and 0.0024 for d 3; Pooled SEM for d 0 is 0.313, 0.204 for d 1, and 0.0024 for d 3; Per day N = 16 and n = 4. Means with different superscripts are considered different (a,b).
Figure Legend Snippet: The comparative effect of 1% Sodium Bisulfate, SBS, and 200 ppm of peracetic acid, PAA, utilized alone or in combination as antimicrobial 15 s part dips on the presence of Salmonella Enteritidis on whole chicken drumsticks on d 0, 1, and 3. In the current study, drumsticks were artificially inoculated with 10 7 CFU/g of S . Enteritidis and subsequently treated in 300 mL of antimicrobial treatments to identify the remaining population of Salmonella . In the study, there were eight treatments, consisting of: a no treatment Control, tap water (TW), tap water with the addition of either 1, 2, or 3% SBS indicated as TW+SBS1, TW+SBS2, and TW+SBS3, and the combination of 1, 2, and 3% SBS with 200 ppm of peracetic acid (PAA), represented as TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA. However, in the current figure only the Control, TW, TW+SBS1, and TW+SBS1+PAA is represented and is separated by d 0, 1, and 3 of 4°C incubation. Individual SEM for Control, TW, TW+SBS1, and TW+SBS1+PAA was 0.293, 0.284, 0.196, and 0.432 for d 0; 0.205, 0.309, 0.150, and 0.086 for d 1; and 0.223, 0.278, 0.064, and 0.078 for d 3, respectively. P -value for d 0 is 0.0116, 0.0067 for d 1, and 0.0024 for d 3; Pooled SEM for d 0 is 0.313, 0.204 for d 1, and 0.0024 for d 3; Per day N = 16 and n = 4. Means with different superscripts are considered different (a,b).

Techniques Used: Incubation

The comparative effect of 2% Sodium Bisulfate, SBS, and 200 ppm of peracetic acid, PAA, utilized alone or in combination as antimicrobial 15 s part dips on the population of Salmonella Enteritidis on whole chicken drumsticks on d 0, 1, and 3. In the current study, drumsticks were artificially inoculated with 10 7 CFU/g of S . Enteritidis and subsequently treated in 300 mL of antimicrobial treatments to identify the remaining population of Salmonella . In the study, there were eight treatments, consisting of: a no treatment Control, tap water (TW), tap water with the addition of either 1, 2, or 3% SBS indicated as TW+SBS1, TW+SBS2, and TW+SBS3, and the combination of 1, 2, and 3% SBS with 200 ppm of peracetic acid (PAA), represented as TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA. However, in the current figure only the Control, TW, TW+SBS2, and TW+SBS2+PAA is represented and is separated by d 0, 1, and 3 of 4°C incubation. Individual SEM for Control, TW, TW+SBS2, and TW+SBS2+PAA was 0.293, 0.284, 0.101, and 0.396 for d 0; 0.205, 0.309, 0.143, and 0.136 for d 1; and 0.223, 0.278, 0.086, and 0.125 for d 3, respectively. P -value for d 0 is 0.0343, 0.0005 for d 1, and
Figure Legend Snippet: The comparative effect of 2% Sodium Bisulfate, SBS, and 200 ppm of peracetic acid, PAA, utilized alone or in combination as antimicrobial 15 s part dips on the population of Salmonella Enteritidis on whole chicken drumsticks on d 0, 1, and 3. In the current study, drumsticks were artificially inoculated with 10 7 CFU/g of S . Enteritidis and subsequently treated in 300 mL of antimicrobial treatments to identify the remaining population of Salmonella . In the study, there were eight treatments, consisting of: a no treatment Control, tap water (TW), tap water with the addition of either 1, 2, or 3% SBS indicated as TW+SBS1, TW+SBS2, and TW+SBS3, and the combination of 1, 2, and 3% SBS with 200 ppm of peracetic acid (PAA), represented as TW+SBS1+PAA, TW+SBS2+PAA, and TW+SBS3+PAA. However, in the current figure only the Control, TW, TW+SBS2, and TW+SBS2+PAA is represented and is separated by d 0, 1, and 3 of 4°C incubation. Individual SEM for Control, TW, TW+SBS2, and TW+SBS2+PAA was 0.293, 0.284, 0.101, and 0.396 for d 0; 0.205, 0.309, 0.143, and 0.136 for d 1; and 0.223, 0.278, 0.086, and 0.125 for d 3, respectively. P -value for d 0 is 0.0343, 0.0005 for d 1, and

Techniques Used: Incubation

21) Product Images from "SiOx-based graphite composite anode and efficient binders: practical applications in lithium-ion batteries †"

Article Title: SiOx-based graphite composite anode and efficient binders: practical applications in lithium-ion batteries †

Journal: RSC Advances

doi: 10.1039/d0ra10283k

(a) CV and (b) EIS of the SiO x –graphite anode coin cells with SBR and PAA binders at pristine and first lithiated states.
Figure Legend Snippet: (a) CV and (b) EIS of the SiO x –graphite anode coin cells with SBR and PAA binders at pristine and first lithiated states.

Techniques Used: Impedance Spectroscopy

(a) Rate performance of coin cells with SBR and PAA binders. The current rates at 0.1–1C (b) action mechanism of the SBR binder.
Figure Legend Snippet: (a) Rate performance of coin cells with SBR and PAA binders. The current rates at 0.1–1C (b) action mechanism of the SBR binder.

Techniques Used:

Cycle performance and battery swelling with SBR and PAA binders.
Figure Legend Snippet: Cycle performance and battery swelling with SBR and PAA binders.

Techniques Used:

Peeling strength of the SiO x –graphite anode electrodes with two binders (a) SBR and (b) PAA.
Figure Legend Snippet: Peeling strength of the SiO x –graphite anode electrodes with two binders (a) SBR and (b) PAA.

Techniques Used:

Cycle performance and capacity retention of coin cells with SBR and PAA binders.
Figure Legend Snippet: Cycle performance and capacity retention of coin cells with SBR and PAA binders.

Techniques Used:

The first two charge and discharge curves of anodes with (a) SBR binder and (b) PAA binder.
Figure Legend Snippet: The first two charge and discharge curves of anodes with (a) SBR binder and (b) PAA binder.

Techniques Used:

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    Millipore sds polyacrylamide gel electrophoresis
    Phenotype detection of the P. multocida Δ crp mutant. a Analysis of the growth curve. The parent strain, S416 (Δ crp ) and S416 (pQK176) were grown in BHI broth or BHI broth supplemented with kanamycin, and the OD 600 values were measured every 2 h over a period of 14 h. The data are expressed as the means ± SD, and the asterisks indicate significant differences compared with the parent strain. b and c Profiles of the OMPs and LPS of P. multocida . OMPs or LPSs were isolated from the parent strain or the Δ crp mutant and then analyzed by <t>SDS-PAGE.</t> Coomassie blue staining and silver staining were then performed to visualize the OMPs ( b ) and LPSs ( c ), respectively. M refers to the protein marker
    Sds Polyacrylamide Gel Electrophoresis, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore sodium dodecyl sulfate polyacrylamide gel electrophoresis sds page
    FgMyo1 is required for toxin biosynthesis. (A) Localization of FgMyo1-RFP in hyphae of PH-1::FgMyo1-RFP growth in toxin non-inducing media PDB and MM at 28°C for 48 h. Bar = 10 μm. (B ) FgMyo1-RFP was co-localized with Tri1-GFP under the toxin inducing condition. Bar = 10 μm. (C) The interaction of FgMyo1 with Tri1 was confirmed by co-immunoprecipitation (Co-IP) analysis. Total proteins (input) extracted from the strain bearing FgMyo1-3×Flag and Tri1-GFP constructs or a single construct (FgMyo1-3×Flag or Tri1-GFP) were subjected to <t>SDS-PAGE,</t> and immunoblots were incubated with anti-Flag and anti-GFP antibodies, as indicated (Input panel). Each protein sample was pulled down using anti-Flag agarose and further detected with anti-GFP antibody (Flag pull-down panel). The protein samples were also incubated with the anti-GAPDH antibody as a reference. (D) The interaction of FgMyo1 with Tri1 was confirmed by bimolecular fluorescence complementation (BiFC) analysis. The constructs of pFgTri1-YFP N and pFgMyo1-YFP C were co-transformed into PH-1 to generate the strain FgTri1-YFP N +FgMyo1-YFP C . The strains bearing a single construct (FgMyo1-YFP C or FgTri1-YFP N ) were used as negative controls. The YFP signals in hyphae of each strain grown in the TBI medium were examined under a confocal microscope. Bar = 10 μm. (E) The sensitivity of FgMyo1 derived mutants towards phenamacril. The wild-type PH-1, FgMyo1 silencing mutant FgMyo1-S2, inducible mutant Pzear-FgMYO1, and the point mutation strain FgMyo1 E420K were incubated on PDA supplemented with 0.3 μg/ml phenamacril (left panel). For the inducible mutant, PDA was also added with (+) or without (-) the inducer 30 μg/ml β-estradiol. Mycelial growth inhibition of each strain by phenamacril was quantified (right panel). Values on the bars followed by the same letter are not significantly different according to a Fisher’s least significant difference (LSD) test at P = 0.05. (F) The toxisome formation patterns in FgMyo1 derived mutants. Each strain was grown in TBI, and images were taken after incubation for 48 h (left-upper panel). The accumulation of Tri1-GFP protein in each strain was determined by western blot assay with the anti-GFP antibody. The protein samples were also incubated with the anti-GAPDH antibody as a reference (left-lower panel). The intensities of GFP signals in each strain were also quantified. Values on the bars followed by the same letter are not significantly different according to a Fisher’s least significant difference (LSD) test at P = 0.05 (right panel). (G) The DON production of FgMyo1 derived mutants. DON was extracted from mycelia of each strain grown in TBI for 7 days. Values on the bars followed by the same letter are not significantly different according to a Fisher’s least significant difference (LSD) test at P = 0.05.
    Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis Sds Page, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Phenotype detection of the P. multocida Δ crp mutant. a Analysis of the growth curve. The parent strain, S416 (Δ crp ) and S416 (pQK176) were grown in BHI broth or BHI broth supplemented with kanamycin, and the OD 600 values were measured every 2 h over a period of 14 h. The data are expressed as the means ± SD, and the asterisks indicate significant differences compared with the parent strain. b and c Profiles of the OMPs and LPS of P. multocida . OMPs or LPSs were isolated from the parent strain or the Δ crp mutant and then analyzed by SDS-PAGE. Coomassie blue staining and silver staining were then performed to visualize the OMPs ( b ) and LPSs ( c ), respectively. M refers to the protein marker

    Journal: BMC Microbiology

    Article Title: Identification of the crp gene in avian Pasteurella multocida and evaluation of the effects of crp deletion on its phenotype, virulence and immunogenicity

    doi: 10.1186/s12866-016-0739-y

    Figure Lengend Snippet: Phenotype detection of the P. multocida Δ crp mutant. a Analysis of the growth curve. The parent strain, S416 (Δ crp ) and S416 (pQK176) were grown in BHI broth or BHI broth supplemented with kanamycin, and the OD 600 values were measured every 2 h over a period of 14 h. The data are expressed as the means ± SD, and the asterisks indicate significant differences compared with the parent strain. b and c Profiles of the OMPs and LPS of P. multocida . OMPs or LPSs were isolated from the parent strain or the Δ crp mutant and then analyzed by SDS-PAGE. Coomassie blue staining and silver staining were then performed to visualize the OMPs ( b ) and LPSs ( c ), respectively. M refers to the protein marker

    Article Snippet: The samples were then subjected to 12.5 % SDS-polyacrylamide gel electrophoresis (PAGE) followed by Coomassie Brilliant Blue R-250 staining (Sigma-Aldrich).

    Techniques: Mutagenesis, Isolation, SDS Page, Staining, Silver Staining, Marker

    FgMyo1 is required for toxin biosynthesis. (A) Localization of FgMyo1-RFP in hyphae of PH-1::FgMyo1-RFP growth in toxin non-inducing media PDB and MM at 28°C for 48 h. Bar = 10 μm. (B ) FgMyo1-RFP was co-localized with Tri1-GFP under the toxin inducing condition. Bar = 10 μm. (C) The interaction of FgMyo1 with Tri1 was confirmed by co-immunoprecipitation (Co-IP) analysis. Total proteins (input) extracted from the strain bearing FgMyo1-3×Flag and Tri1-GFP constructs or a single construct (FgMyo1-3×Flag or Tri1-GFP) were subjected to SDS-PAGE, and immunoblots were incubated with anti-Flag and anti-GFP antibodies, as indicated (Input panel). Each protein sample was pulled down using anti-Flag agarose and further detected with anti-GFP antibody (Flag pull-down panel). The protein samples were also incubated with the anti-GAPDH antibody as a reference. (D) The interaction of FgMyo1 with Tri1 was confirmed by bimolecular fluorescence complementation (BiFC) analysis. The constructs of pFgTri1-YFP N and pFgMyo1-YFP C were co-transformed into PH-1 to generate the strain FgTri1-YFP N +FgMyo1-YFP C . The strains bearing a single construct (FgMyo1-YFP C or FgTri1-YFP N ) were used as negative controls. The YFP signals in hyphae of each strain grown in the TBI medium were examined under a confocal microscope. Bar = 10 μm. (E) The sensitivity of FgMyo1 derived mutants towards phenamacril. The wild-type PH-1, FgMyo1 silencing mutant FgMyo1-S2, inducible mutant Pzear-FgMYO1, and the point mutation strain FgMyo1 E420K were incubated on PDA supplemented with 0.3 μg/ml phenamacril (left panel). For the inducible mutant, PDA was also added with (+) or without (-) the inducer 30 μg/ml β-estradiol. Mycelial growth inhibition of each strain by phenamacril was quantified (right panel). Values on the bars followed by the same letter are not significantly different according to a Fisher’s least significant difference (LSD) test at P = 0.05. (F) The toxisome formation patterns in FgMyo1 derived mutants. Each strain was grown in TBI, and images were taken after incubation for 48 h (left-upper panel). The accumulation of Tri1-GFP protein in each strain was determined by western blot assay with the anti-GFP antibody. The protein samples were also incubated with the anti-GAPDH antibody as a reference (left-lower panel). The intensities of GFP signals in each strain were also quantified. Values on the bars followed by the same letter are not significantly different according to a Fisher’s least significant difference (LSD) test at P = 0.05 (right panel). (G) The DON production of FgMyo1 derived mutants. DON was extracted from mycelia of each strain grown in TBI for 7 days. Values on the bars followed by the same letter are not significantly different according to a Fisher’s least significant difference (LSD) test at P = 0.05.

    Journal: PLoS Pathogens

    Article Title: The fungal myosin I is essential for Fusarium toxisome formation

    doi: 10.1371/journal.ppat.1006827

    Figure Lengend Snippet: FgMyo1 is required for toxin biosynthesis. (A) Localization of FgMyo1-RFP in hyphae of PH-1::FgMyo1-RFP growth in toxin non-inducing media PDB and MM at 28°C for 48 h. Bar = 10 μm. (B ) FgMyo1-RFP was co-localized with Tri1-GFP under the toxin inducing condition. Bar = 10 μm. (C) The interaction of FgMyo1 with Tri1 was confirmed by co-immunoprecipitation (Co-IP) analysis. Total proteins (input) extracted from the strain bearing FgMyo1-3×Flag and Tri1-GFP constructs or a single construct (FgMyo1-3×Flag or Tri1-GFP) were subjected to SDS-PAGE, and immunoblots were incubated with anti-Flag and anti-GFP antibodies, as indicated (Input panel). Each protein sample was pulled down using anti-Flag agarose and further detected with anti-GFP antibody (Flag pull-down panel). The protein samples were also incubated with the anti-GAPDH antibody as a reference. (D) The interaction of FgMyo1 with Tri1 was confirmed by bimolecular fluorescence complementation (BiFC) analysis. The constructs of pFgTri1-YFP N and pFgMyo1-YFP C were co-transformed into PH-1 to generate the strain FgTri1-YFP N +FgMyo1-YFP C . The strains bearing a single construct (FgMyo1-YFP C or FgTri1-YFP N ) were used as negative controls. The YFP signals in hyphae of each strain grown in the TBI medium were examined under a confocal microscope. Bar = 10 μm. (E) The sensitivity of FgMyo1 derived mutants towards phenamacril. The wild-type PH-1, FgMyo1 silencing mutant FgMyo1-S2, inducible mutant Pzear-FgMYO1, and the point mutation strain FgMyo1 E420K were incubated on PDA supplemented with 0.3 μg/ml phenamacril (left panel). For the inducible mutant, PDA was also added with (+) or without (-) the inducer 30 μg/ml β-estradiol. Mycelial growth inhibition of each strain by phenamacril was quantified (right panel). Values on the bars followed by the same letter are not significantly different according to a Fisher’s least significant difference (LSD) test at P = 0.05. (F) The toxisome formation patterns in FgMyo1 derived mutants. Each strain was grown in TBI, and images were taken after incubation for 48 h (left-upper panel). The accumulation of Tri1-GFP protein in each strain was determined by western blot assay with the anti-GFP antibody. The protein samples were also incubated with the anti-GAPDH antibody as a reference (left-lower panel). The intensities of GFP signals in each strain were also quantified. Values on the bars followed by the same letter are not significantly different according to a Fisher’s least significant difference (LSD) test at P = 0.05 (right panel). (G) The DON production of FgMyo1 derived mutants. DON was extracted from mycelia of each strain grown in TBI for 7 days. Values on the bars followed by the same letter are not significantly different according to a Fisher’s least significant difference (LSD) test at P = 0.05.

    Article Snippet: The resulting proteins were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to Immobilon-P transfer membrane (Millipore, Billerica, MA, USA).

    Techniques: Immunoprecipitation, Co-Immunoprecipitation Assay, Construct, SDS Page, Western Blot, Incubation, Fluorescence, Bimolecular Fluorescence Complementation Assay, Transformation Assay, Microscopy, Derivative Assay, Mutagenesis, Inhibition

    The actin cytoskeleton is involved in toxisome formation. (A) The interaction of Actin-RFP and Tri1-GFP was verified by the Co-IP assay. Total proteins (input) extracted from the strain bearing Actin-RFP and Tri1-GFP constructs or a single construct (Actin-RFP or Tri1-GFP) were subjected to SDS-PAGE, and immunoblots were incubated with anti-GFP and anti-RFP antibodies, as indicated (Input panel). Each protein sample was pulled down using anti-GFP agarose and further detected with anti-RFP antibody (GFP pull-down panel). The protein samples were also incubated with the anti-GAPDH antibody as a reference. (B) Co-IP analysis for verification of the interaction between FgMyo1-GFP and Actin-RFP. Total proteins (input) extracted from the strain bearing Actin-RFP and FgMyo1-GFP constructs or a single construct (Actin-RFP or FgMyo1-GFP) were subjected to SDS-PAGE, and immunoblots were incubated with anti-Flag and anti-GFP antibodies, as indicated (Input panel). Each protein sample was pulled down using anti-GFP agarose and further detected with anti-RFP antibody (GFP pull-down panel). The protein samples were also incubated with the anti-GAPDH antibody as a reference. (C) The actin polymerization inhibitor latrunculin A inhibited toxisome formation. After growth in TBI for 24 h, ΔTri1::Tri1-GFP was treated with 0.1 μg/ml latrunculin A for another 24 h before examination (left panel). The solvent DMSO was used as a control. Bar = 10 μm. The accumulation of Tri1-GFP protein was further verified by western blotting assay using the anti-GFP antibody (right panel). The protein samples were also incubated with the anti-GAPDH antibody as a reference. (D) DON was extracted from mycelia of PH-1 grown in TBI supplemented with 0.1 μg/ml latrunculin A. The solvent DMSO was used as a control. Values on the bars followed by different letters are significantly different according to a Fisher’s least significant difference (LSD) test at P = 0.05.

    Journal: PLoS Pathogens

    Article Title: The fungal myosin I is essential for Fusarium toxisome formation

    doi: 10.1371/journal.ppat.1006827

    Figure Lengend Snippet: The actin cytoskeleton is involved in toxisome formation. (A) The interaction of Actin-RFP and Tri1-GFP was verified by the Co-IP assay. Total proteins (input) extracted from the strain bearing Actin-RFP and Tri1-GFP constructs or a single construct (Actin-RFP or Tri1-GFP) were subjected to SDS-PAGE, and immunoblots were incubated with anti-GFP and anti-RFP antibodies, as indicated (Input panel). Each protein sample was pulled down using anti-GFP agarose and further detected with anti-RFP antibody (GFP pull-down panel). The protein samples were also incubated with the anti-GAPDH antibody as a reference. (B) Co-IP analysis for verification of the interaction between FgMyo1-GFP and Actin-RFP. Total proteins (input) extracted from the strain bearing Actin-RFP and FgMyo1-GFP constructs or a single construct (Actin-RFP or FgMyo1-GFP) were subjected to SDS-PAGE, and immunoblots were incubated with anti-Flag and anti-GFP antibodies, as indicated (Input panel). Each protein sample was pulled down using anti-GFP agarose and further detected with anti-RFP antibody (GFP pull-down panel). The protein samples were also incubated with the anti-GAPDH antibody as a reference. (C) The actin polymerization inhibitor latrunculin A inhibited toxisome formation. After growth in TBI for 24 h, ΔTri1::Tri1-GFP was treated with 0.1 μg/ml latrunculin A for another 24 h before examination (left panel). The solvent DMSO was used as a control. Bar = 10 μm. The accumulation of Tri1-GFP protein was further verified by western blotting assay using the anti-GFP antibody (right panel). The protein samples were also incubated with the anti-GAPDH antibody as a reference. (D) DON was extracted from mycelia of PH-1 grown in TBI supplemented with 0.1 μg/ml latrunculin A. The solvent DMSO was used as a control. Values on the bars followed by different letters are significantly different according to a Fisher’s least significant difference (LSD) test at P = 0.05.

    Article Snippet: The resulting proteins were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to Immobilon-P transfer membrane (Millipore, Billerica, MA, USA).

    Techniques: Co-Immunoprecipitation Assay, Construct, SDS Page, Western Blot, Incubation