phospho foxo  (Cell Signaling Technology Inc)


Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc phospho foxo
    17-AAG induces downregulation of <t>FOXO</t> and ERK1/2 signaling mediators in bladder cancer cells. Detection of total and constitutively phosphorylated protein levels of FOXO transcription factors and <t>Erk1/2</t> <t>(p44/42)</t> kinases in all three bladder cancer cell lines, upon 17-AAG administration for 24 hours, via a Western blotting approach. All immunoblotting experiments were repeated three times.
    Phospho Foxo, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho foxo/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho foxo - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "17-Allylamino-17-demethoxygeldanamycin induces downregulation of critical Hsp90 protein clients and results in cell cycle arrest and apoptosis of human urinary bladder cancer cells"

    Article Title: 17-Allylamino-17-demethoxygeldanamycin induces downregulation of critical Hsp90 protein clients and results in cell cycle arrest and apoptosis of human urinary bladder cancer cells

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-10-481

    17-AAG induces downregulation of FOXO and ERK1/2 signaling mediators in bladder cancer cells. Detection of total and constitutively phosphorylated protein levels of FOXO transcription factors and Erk1/2 (p44/42) kinases in all three bladder cancer cell lines, upon 17-AAG administration for 24 hours, via a Western blotting approach. All immunoblotting experiments were repeated three times.
    Figure Legend Snippet: 17-AAG induces downregulation of FOXO and ERK1/2 signaling mediators in bladder cancer cells. Detection of total and constitutively phosphorylated protein levels of FOXO transcription factors and Erk1/2 (p44/42) kinases in all three bladder cancer cell lines, upon 17-AAG administration for 24 hours, via a Western blotting approach. All immunoblotting experiments were repeated three times.

    Techniques Used: Western Blot

    phospho foxo  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc phospho foxo
    17-AAG induces downregulation of <t>FOXO</t> and ERK1/2 signaling mediators in bladder cancer cells. Detection of total and constitutively phosphorylated protein levels of FOXO transcription factors and <t>Erk1/2</t> <t>(p44/42)</t> kinases in all three bladder cancer cell lines, upon 17-AAG administration for 24 hours, via a Western blotting approach. All immunoblotting experiments were repeated three times.
    Phospho Foxo, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho foxo/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho foxo - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "17-Allylamino-17-demethoxygeldanamycin induces downregulation of critical Hsp90 protein clients and results in cell cycle arrest and apoptosis of human urinary bladder cancer cells"

    Article Title: 17-Allylamino-17-demethoxygeldanamycin induces downregulation of critical Hsp90 protein clients and results in cell cycle arrest and apoptosis of human urinary bladder cancer cells

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-10-481

    17-AAG induces downregulation of FOXO and ERK1/2 signaling mediators in bladder cancer cells. Detection of total and constitutively phosphorylated protein levels of FOXO transcription factors and Erk1/2 (p44/42) kinases in all three bladder cancer cell lines, upon 17-AAG administration for 24 hours, via a Western blotting approach. All immunoblotting experiments were repeated three times.
    Figure Legend Snippet: 17-AAG induces downregulation of FOXO and ERK1/2 signaling mediators in bladder cancer cells. Detection of total and constitutively phosphorylated protein levels of FOXO transcription factors and Erk1/2 (p44/42) kinases in all three bladder cancer cell lines, upon 17-AAG administration for 24 hours, via a Western blotting approach. All immunoblotting experiments were repeated three times.

    Techniques Used: Western Blot

    mouse anti hiv 1 p24 monoclonal antibody  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc mouse anti hiv 1 p24 monoclonal antibody
    Virological features of BA.2 in vitro (A) Scheme for the chimeric recombinant SARS-CoV-2 used in this study. The SARS-CoV-2 genome and its genes are shown. The template was SARS-CoV-2 strain WK-521 (PANGO lineage A, GISAID ID: EPI_ISL_408667), and the S genes were swapped with those of the respective lineages/strains (GISAID IDs are indicated in the figure). ORF7a was swapped with the sfGFP gene. (B) Growth kinetics of chimeric recombinant SARS-CoV-2 in Vero cells, VeroE6/TMPRSS2 cells, Calu-3 cells, and human nasal epithelial cells. (C) Fluorescence microscopy. The GFP area was measured in infected VeroE6/TMPRSS2 cells (multiplicity of infection [m.o.i.] 0.01) at 48 h.p.i. Left, representative panels. Higher-magnification views of the regions indicated by squares are shown at the bottom. Representative time-course data are shown in <xref ref-type=Figure S2 E. Scale bars, 500 μm. Right, the summarized results. The numbers in the panel indicate the numbers of GFP-positive cells counted. (D) Plaque assay. Representative panels (left) and a summary of the recorded plaque diameters (20 plaques per virus) (right) are shown. (E) S expression on the cell surface. Representative histograms stained with an anti-S1/S2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates mean fluorescence intensity (MFI). Gray histograms indicate isotype controls. (F) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (G) Western blotting. Left, representative blots of S-expressing cells (top) and pseudovirus (bottom). ACTB is an internal control for the cells, whereas HIV-1 p24 is an internal control for the pseudovirus. kDa, kilodalton. Middle, the ratio of S2 to the full-length S plus S2 proteins in the cells. Right, the ratio of S2 to HIV-1 p24 in the pseudovirus (supernatant). (H) Pseudovirus assay. The percent infectivity compared with that of the virus pseudotyped with B.1.1 S are shown. (I) Binding affinity of SARS-CoV-2 S RBD to ACE2 by yeast surface display. The percentage of SARS-CoV-2 S RBD expressed on yeast binding to soluble ACE2 (left) and the summarized K D values (right) are shown. (J) TMPRSS2 expression on the cell surface. Representative histograms stained with an anti-TMPRSS2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates MFI. Gray histograms indicate the isotype controls. (K) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (L) Fold increase in pseudovirus infectivity based on TMPRSS2 expression. (M) E64d treatment. IC50, 50% inhibitory concentration; ND, not determined. (N) Growth kinetics of chimeric recombinant SARS-CoV-2 in HK293-ACE2 and HEK293-ACE2/TMPRSS2 cells. Assays were performed in quadruplicate (B, H, L, J, and N), octuplicate (B, most right) or triplicate (E–G, I, J, K, and M), and the presented data are expressed as the average ± SD. Each dot indicates the result of an individual plaque (D) and an individual replicate (E, G– J, L and I). Statistically significant differences between BA.2 and other variants across time points were determined by multiple regression (B, F, K, and N). Family-wise error rates (FWERs) calculated using the Holm method are indicated in the figures. Statistically significant differences between BA.1 and BA.2 were determined by two-sided Mann-Whitney U tests (C and D), two-sided Student’s t tests (E, H, and I), or two-sided paired t test (G). See also Figure S3 . " width="250" height="auto" />
    Mouse Anti Hiv 1 P24 Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti hiv 1 p24 monoclonal antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti hiv 1 p24 monoclonal antibody - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Virological characteristics of the SARS-CoV-2 Omicron BA.2 spike"

    Article Title: Virological characteristics of the SARS-CoV-2 Omicron BA.2 spike

    Journal: Cell

    doi: 10.1016/j.cell.2022.04.035

    Virological features of BA.2 in vitro (A) Scheme for the chimeric recombinant SARS-CoV-2 used in this study. The SARS-CoV-2 genome and its genes are shown. The template was SARS-CoV-2 strain WK-521 (PANGO lineage A, GISAID ID: EPI_ISL_408667), and the S genes were swapped with those of the respective lineages/strains (GISAID IDs are indicated in the figure). ORF7a was swapped with the sfGFP gene. (B) Growth kinetics of chimeric recombinant SARS-CoV-2 in Vero cells, VeroE6/TMPRSS2 cells, Calu-3 cells, and human nasal epithelial cells. (C) Fluorescence microscopy. The GFP area was measured in infected VeroE6/TMPRSS2 cells (multiplicity of infection [m.o.i.] 0.01) at 48 h.p.i. Left, representative panels. Higher-magnification views of the regions indicated by squares are shown at the bottom. Representative time-course data are shown in <xref ref-type=Figure S2 E. Scale bars, 500 μm. Right, the summarized results. The numbers in the panel indicate the numbers of GFP-positive cells counted. (D) Plaque assay. Representative panels (left) and a summary of the recorded plaque diameters (20 plaques per virus) (right) are shown. (E) S expression on the cell surface. Representative histograms stained with an anti-S1/S2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates mean fluorescence intensity (MFI). Gray histograms indicate isotype controls. (F) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (G) Western blotting. Left, representative blots of S-expressing cells (top) and pseudovirus (bottom). ACTB is an internal control for the cells, whereas HIV-1 p24 is an internal control for the pseudovirus. kDa, kilodalton. Middle, the ratio of S2 to the full-length S plus S2 proteins in the cells. Right, the ratio of S2 to HIV-1 p24 in the pseudovirus (supernatant). (H) Pseudovirus assay. The percent infectivity compared with that of the virus pseudotyped with B.1.1 S are shown. (I) Binding affinity of SARS-CoV-2 S RBD to ACE2 by yeast surface display. The percentage of SARS-CoV-2 S RBD expressed on yeast binding to soluble ACE2 (left) and the summarized K D values (right) are shown. (J) TMPRSS2 expression on the cell surface. Representative histograms stained with an anti-TMPRSS2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates MFI. Gray histograms indicate the isotype controls. (K) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (L) Fold increase in pseudovirus infectivity based on TMPRSS2 expression. (M) E64d treatment. IC50, 50% inhibitory concentration; ND, not determined. (N) Growth kinetics of chimeric recombinant SARS-CoV-2 in HK293-ACE2 and HEK293-ACE2/TMPRSS2 cells. Assays were performed in quadruplicate (B, H, L, J, and N), octuplicate (B, most right) or triplicate (E–G, I, J, K, and M), and the presented data are expressed as the average ± SD. Each dot indicates the result of an individual plaque (D) and an individual replicate (E, G– J, L and I). Statistically significant differences between BA.2 and other variants across time points were determined by multiple regression (B, F, K, and N). Family-wise error rates (FWERs) calculated using the Holm method are indicated in the figures. Statistically significant differences between BA.1 and BA.2 were determined by two-sided Mann-Whitney U tests (C and D), two-sided Student’s t tests (E, H, and I), or two-sided paired t test (G). See also Figure S3 . " title="... is an internal control for the cells, whereas HIV-1 p24 is an internal control for the pseudovirus. ..." property="contentUrl" width="100%" height="100%"/>
    Figure Legend Snippet: Virological features of BA.2 in vitro (A) Scheme for the chimeric recombinant SARS-CoV-2 used in this study. The SARS-CoV-2 genome and its genes are shown. The template was SARS-CoV-2 strain WK-521 (PANGO lineage A, GISAID ID: EPI_ISL_408667), and the S genes were swapped with those of the respective lineages/strains (GISAID IDs are indicated in the figure). ORF7a was swapped with the sfGFP gene. (B) Growth kinetics of chimeric recombinant SARS-CoV-2 in Vero cells, VeroE6/TMPRSS2 cells, Calu-3 cells, and human nasal epithelial cells. (C) Fluorescence microscopy. The GFP area was measured in infected VeroE6/TMPRSS2 cells (multiplicity of infection [m.o.i.] 0.01) at 48 h.p.i. Left, representative panels. Higher-magnification views of the regions indicated by squares are shown at the bottom. Representative time-course data are shown in Figure S2 E. Scale bars, 500 μm. Right, the summarized results. The numbers in the panel indicate the numbers of GFP-positive cells counted. (D) Plaque assay. Representative panels (left) and a summary of the recorded plaque diameters (20 plaques per virus) (right) are shown. (E) S expression on the cell surface. Representative histograms stained with an anti-S1/S2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates mean fluorescence intensity (MFI). Gray histograms indicate isotype controls. (F) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (G) Western blotting. Left, representative blots of S-expressing cells (top) and pseudovirus (bottom). ACTB is an internal control for the cells, whereas HIV-1 p24 is an internal control for the pseudovirus. kDa, kilodalton. Middle, the ratio of S2 to the full-length S plus S2 proteins in the cells. Right, the ratio of S2 to HIV-1 p24 in the pseudovirus (supernatant). (H) Pseudovirus assay. The percent infectivity compared with that of the virus pseudotyped with B.1.1 S are shown. (I) Binding affinity of SARS-CoV-2 S RBD to ACE2 by yeast surface display. The percentage of SARS-CoV-2 S RBD expressed on yeast binding to soluble ACE2 (left) and the summarized K D values (right) are shown. (J) TMPRSS2 expression on the cell surface. Representative histograms stained with an anti-TMPRSS2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates MFI. Gray histograms indicate the isotype controls. (K) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (L) Fold increase in pseudovirus infectivity based on TMPRSS2 expression. (M) E64d treatment. IC50, 50% inhibitory concentration; ND, not determined. (N) Growth kinetics of chimeric recombinant SARS-CoV-2 in HK293-ACE2 and HEK293-ACE2/TMPRSS2 cells. Assays were performed in quadruplicate (B, H, L, J, and N), octuplicate (B, most right) or triplicate (E–G, I, J, K, and M), and the presented data are expressed as the average ± SD. Each dot indicates the result of an individual plaque (D) and an individual replicate (E, G– J, L and I). Statistically significant differences between BA.2 and other variants across time points were determined by multiple regression (B, F, K, and N). Family-wise error rates (FWERs) calculated using the Holm method are indicated in the figures. Statistically significant differences between BA.1 and BA.2 were determined by two-sided Mann-Whitney U tests (C and D), two-sided Student’s t tests (E, H, and I), or two-sided paired t test (G). See also Figure S3 .

    Techniques Used: In Vitro, Recombinant, Fluorescence, Microscopy, Infection, Plaque Assay, Expressing, Staining, Single Vesicle Fusion Assay, Activity Assay, Western Blot, Binding Assay, Concentration Assay, MANN-WHITNEY


    Figure Legend Snippet:

    Techniques Used: Recombinant, Expressing, Protease Inhibitor, Activity Assay, Luciferase, CCK-8 Assay, Sequencing, Plasmid Preparation, Software

    mouse anti hiv 1 p24 monoclonal antibody  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc mouse anti hiv 1 p24 monoclonal antibody
    ( A ) Scheme of S chimeras used in this study. The numbers in parentheses are identical to those in – and . NTD, N-terminal domain; RBD, receptor-binding domain; TMD, transmembrane domain. ( B ) Pseudovirus assay. <t>HIV-1-based</t> reporter viruses pseudotyped with SARS-CoV-2 S chimeras (summarized in ) were prepared. The pseudoviruses were inoculated into HOS-ACE2/TMPRSS2 cells at 1,000 ng HIV-1 <t>p24</t> antigen, and the percentages of infectivity compared to that of the virus pseudotyped with B.1 S (spike 1) are shown. ( C and D ) Western blot. Representative blots of S-expressing cells and supernatants ( C ) and quantified band intensity (the ratio of S2 to the full-length S plus S2 proteins for “cell”; the ratio of S2 to HIV-1 p24 for “supernatant”) ( D ) are shown. M, mock (empty vector-transfected). ( E ) Flow cytometry. The summarized results of the surface S expression are shown. MFI, mean fluorescent intensity; M, mock (empty vector-transfected). ( F ) SARS-CoV-2 S-based fusion assay. The fusion activity was measured as described in the STAR⍰METHODS, and fusion activity (arbitrary units) is shown. For the target cells, HEK293 cells expressing ACE2 and TMPRSS2 (filled) and HEK293 cells expressing ACE2 (open) were used. The results for B.1 S or Omicron S are shown in other panels as black and green lines, respectively. The results in HEK293-ACE2/TMPRSS2 cells and HEK293-ACE2 cells are shown as normal or broken lines, respectively. ( G ) Scheme of the S-chimeric recombinant SARS-CoV-2 used in this study. FCS, furin cleavage site. The backbone is SARS-CoV-2 strain WK-521 (GISAID ID: EPI_ISL_408667, A lineage) ( Torii et al ., 2021 ). Note that the ORF7a gene is swapped with the sfGFP gene. The numbers in parentheses are identical to those in – . ( H-J ) SARS-CoV-2 infection. VeroE6/TMPRSS2 cells were infected with a series of chimeric recombinant SARS-CoV-2 (shown in G ) at multiplicity of infection (m.o.i.) 0.01. Viral RNA in the supernatant ( H ) and GFP intensity ( I ) were measured using routine techniques. The result for Omicron (virus II) is shown in other panels as a broken green line. ( J ) Syncytium formation. Left, GFP-positive area at 48 h.p.i. Scale bar, 500 μm. Right, summarized results. I, n=6,483 cells; II, n=5,393 cells; III, n=8,704 cells; IV, n=13,188 cells; and V, n=12,749 cells. Representative images are shown in . ( K ) Plaque assay. Left, representative figures. Right, summary of the plaque diameters (20 plaques per virus). Data are expressed as the mean with SD ( B, D-F, H and K ) or the median with 95% confidence interval (CI) ( J ). Assays were performed in quadruplicate ( B, H ) or triplicate ( D-F ). Each dot indicates the result of an individual replicate ( B, D and E ) or an individual plaque ( K ). Statistically significant differences (*P<0.05) versus Omicron S (pseudovirus 2 for B, D and E , virus II for J and K ) were determined by two-sided Student’s t test ( B and E ), two-sided paired t test ( D ), or two-sided Mann–Whitney U test ( J and K ). In F, H and I , statistically significant differences [*familywise error rates (FWERs)<0.05] versus Omicron (spike 2 or virus II) (except for the rightmost panel in F ) or B.1 (spike 1 or virus I) (rightmost panel in F ) through timepoints were determined by multiple regression. FWERs were calculated using the Holm method. See also .
    Mouse Anti Hiv 1 P24 Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti hiv 1 p24 monoclonal antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti hiv 1 p24 monoclonal antibody - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "SARS-CoV-2 spike S375F mutation characterizes the Omicron BA.1 variant"

    Article Title: SARS-CoV-2 spike S375F mutation characterizes the Omicron BA.1 variant

    Journal: bioRxiv

    doi: 10.1101/2022.04.03.486864

    ( A ) Scheme of S chimeras used in this study. The numbers in parentheses are identical to those in – and . NTD, N-terminal domain; RBD, receptor-binding domain; TMD, transmembrane domain. ( B ) Pseudovirus assay. HIV-1-based reporter viruses pseudotyped with SARS-CoV-2 S chimeras (summarized in ) were prepared. The pseudoviruses were inoculated into HOS-ACE2/TMPRSS2 cells at 1,000 ng HIV-1 p24 antigen, and the percentages of infectivity compared to that of the virus pseudotyped with B.1 S (spike 1) are shown. ( C and D ) Western blot. Representative blots of S-expressing cells and supernatants ( C ) and quantified band intensity (the ratio of S2 to the full-length S plus S2 proteins for “cell”; the ratio of S2 to HIV-1 p24 for “supernatant”) ( D ) are shown. M, mock (empty vector-transfected). ( E ) Flow cytometry. The summarized results of the surface S expression are shown. MFI, mean fluorescent intensity; M, mock (empty vector-transfected). ( F ) SARS-CoV-2 S-based fusion assay. The fusion activity was measured as described in the STAR⍰METHODS, and fusion activity (arbitrary units) is shown. For the target cells, HEK293 cells expressing ACE2 and TMPRSS2 (filled) and HEK293 cells expressing ACE2 (open) were used. The results for B.1 S or Omicron S are shown in other panels as black and green lines, respectively. The results in HEK293-ACE2/TMPRSS2 cells and HEK293-ACE2 cells are shown as normal or broken lines, respectively. ( G ) Scheme of the S-chimeric recombinant SARS-CoV-2 used in this study. FCS, furin cleavage site. The backbone is SARS-CoV-2 strain WK-521 (GISAID ID: EPI_ISL_408667, A lineage) ( Torii et al ., 2021 ). Note that the ORF7a gene is swapped with the sfGFP gene. The numbers in parentheses are identical to those in – . ( H-J ) SARS-CoV-2 infection. VeroE6/TMPRSS2 cells were infected with a series of chimeric recombinant SARS-CoV-2 (shown in G ) at multiplicity of infection (m.o.i.) 0.01. Viral RNA in the supernatant ( H ) and GFP intensity ( I ) were measured using routine techniques. The result for Omicron (virus II) is shown in other panels as a broken green line. ( J ) Syncytium formation. Left, GFP-positive area at 48 h.p.i. Scale bar, 500 μm. Right, summarized results. I, n=6,483 cells; II, n=5,393 cells; III, n=8,704 cells; IV, n=13,188 cells; and V, n=12,749 cells. Representative images are shown in . ( K ) Plaque assay. Left, representative figures. Right, summary of the plaque diameters (20 plaques per virus). Data are expressed as the mean with SD ( B, D-F, H and K ) or the median with 95% confidence interval (CI) ( J ). Assays were performed in quadruplicate ( B, H ) or triplicate ( D-F ). Each dot indicates the result of an individual replicate ( B, D and E ) or an individual plaque ( K ). Statistically significant differences (*P<0.05) versus Omicron S (pseudovirus 2 for B, D and E , virus II for J and K ) were determined by two-sided Student’s t test ( B and E ), two-sided paired t test ( D ), or two-sided Mann–Whitney U test ( J and K ). In F, H and I , statistically significant differences [*familywise error rates (FWERs)<0.05] versus Omicron (spike 2 or virus II) (except for the rightmost panel in F ) or B.1 (spike 1 or virus I) (rightmost panel in F ) through timepoints were determined by multiple regression. FWERs were calculated using the Holm method. See also .
    Figure Legend Snippet: ( A ) Scheme of S chimeras used in this study. The numbers in parentheses are identical to those in – and . NTD, N-terminal domain; RBD, receptor-binding domain; TMD, transmembrane domain. ( B ) Pseudovirus assay. HIV-1-based reporter viruses pseudotyped with SARS-CoV-2 S chimeras (summarized in ) were prepared. The pseudoviruses were inoculated into HOS-ACE2/TMPRSS2 cells at 1,000 ng HIV-1 p24 antigen, and the percentages of infectivity compared to that of the virus pseudotyped with B.1 S (spike 1) are shown. ( C and D ) Western blot. Representative blots of S-expressing cells and supernatants ( C ) and quantified band intensity (the ratio of S2 to the full-length S plus S2 proteins for “cell”; the ratio of S2 to HIV-1 p24 for “supernatant”) ( D ) are shown. M, mock (empty vector-transfected). ( E ) Flow cytometry. The summarized results of the surface S expression are shown. MFI, mean fluorescent intensity; M, mock (empty vector-transfected). ( F ) SARS-CoV-2 S-based fusion assay. The fusion activity was measured as described in the STAR⍰METHODS, and fusion activity (arbitrary units) is shown. For the target cells, HEK293 cells expressing ACE2 and TMPRSS2 (filled) and HEK293 cells expressing ACE2 (open) were used. The results for B.1 S or Omicron S are shown in other panels as black and green lines, respectively. The results in HEK293-ACE2/TMPRSS2 cells and HEK293-ACE2 cells are shown as normal or broken lines, respectively. ( G ) Scheme of the S-chimeric recombinant SARS-CoV-2 used in this study. FCS, furin cleavage site. The backbone is SARS-CoV-2 strain WK-521 (GISAID ID: EPI_ISL_408667, A lineage) ( Torii et al ., 2021 ). Note that the ORF7a gene is swapped with the sfGFP gene. The numbers in parentheses are identical to those in – . ( H-J ) SARS-CoV-2 infection. VeroE6/TMPRSS2 cells were infected with a series of chimeric recombinant SARS-CoV-2 (shown in G ) at multiplicity of infection (m.o.i.) 0.01. Viral RNA in the supernatant ( H ) and GFP intensity ( I ) were measured using routine techniques. The result for Omicron (virus II) is shown in other panels as a broken green line. ( J ) Syncytium formation. Left, GFP-positive area at 48 h.p.i. Scale bar, 500 μm. Right, summarized results. I, n=6,483 cells; II, n=5,393 cells; III, n=8,704 cells; IV, n=13,188 cells; and V, n=12,749 cells. Representative images are shown in . ( K ) Plaque assay. Left, representative figures. Right, summary of the plaque diameters (20 plaques per virus). Data are expressed as the mean with SD ( B, D-F, H and K ) or the median with 95% confidence interval (CI) ( J ). Assays were performed in quadruplicate ( B, H ) or triplicate ( D-F ). Each dot indicates the result of an individual replicate ( B, D and E ) or an individual plaque ( K ). Statistically significant differences (*P<0.05) versus Omicron S (pseudovirus 2 for B, D and E , virus II for J and K ) were determined by two-sided Student’s t test ( B and E ), two-sided paired t test ( D ), or two-sided Mann–Whitney U test ( J and K ). In F, H and I , statistically significant differences [*familywise error rates (FWERs)<0.05] versus Omicron (spike 2 or virus II) (except for the rightmost panel in F ) or B.1 (spike 1 or virus I) (rightmost panel in F ) through timepoints were determined by multiple regression. FWERs were calculated using the Holm method. See also .

    Techniques Used: Binding Assay, Infection, Western Blot, Expressing, Plasmid Preparation, Transfection, Flow Cytometry, Single Vesicle Fusion Assay, Activity Assay, Recombinant, Plaque Assay, MANN-WHITNEY

    ( A ) Scheme of the S mutants used in this study. The numbers in parentheses are identical to those in – and . ( B ) Pseudovirus assay. HIV-1-based reporter viruses pseudotyped with SARS-CoV-2 S mutants (summarized in A ) were prepared. The pseudoviruses were inoculated into HOS-ACE2/TMPRSS2 cells at 1,000 ng HIV-1 p24 antigen, and the percent infectivity compared to that of the virus pseudotyped with Omicron S (spike 2, top) or B.1 S (spike 1, bottom) are shown. ( C and D ) Western blot. Representative blots of S-expressing cells and supernatants ( C ) and quantified band intensity (the ratio of S2 to the full-length S plus S2 proteins for “cell”; the ratio of S2 to HIV-1 p24 for “supernatant”) ( D ) are shown. M, mock (empty vector-transfected). ( E ) Flow cytometry. The summarized results of the surface S expression are shown. ( F ) SARS-CoV-2 S-based fusion assay. The fusion activity was measured as described in STAR⍰METHODS, and fusion activity (arbitrary units) is shown. For the target cells, HEK293 cells expressing ACE2 and TMPRSS2 (filled) and HEK293 cells expressing ACE2 (open) were used. The results for Omicron S (top) or B.1 S (bottom) are shown in other panels as green and black lines, respectively. The results in HEK293-ACE2/TMPRSS2 cells and HEK293-ACE2 cells are shown as normal and broken lines, respectively. Data are expressed as the mean with SD. Assays were performed in quadruplicate ( B ) or triplicate ( D-F ). In B, D and E , each dot indicates the result of an individual replicate. Statistically significant differences (*P<0.05) versus the respective parental S [Omicron S (pseudovirus 2, top panels) or B.1 S (spike 1, bottom panels)] were determined by two-sided Student’s t test ( B and E ) or two-sided paired t test ( D ). In F , statistically significant differences (*FWERs<0.05) versus the respective parental S [Omicron S (spike 2, top panels) or B.1 S (spike 1, bottom panels)] through timepoints were determined by multiple regression. FWERs were calculated using the Holm method. See also .
    Figure Legend Snippet: ( A ) Scheme of the S mutants used in this study. The numbers in parentheses are identical to those in – and . ( B ) Pseudovirus assay. HIV-1-based reporter viruses pseudotyped with SARS-CoV-2 S mutants (summarized in A ) were prepared. The pseudoviruses were inoculated into HOS-ACE2/TMPRSS2 cells at 1,000 ng HIV-1 p24 antigen, and the percent infectivity compared to that of the virus pseudotyped with Omicron S (spike 2, top) or B.1 S (spike 1, bottom) are shown. ( C and D ) Western blot. Representative blots of S-expressing cells and supernatants ( C ) and quantified band intensity (the ratio of S2 to the full-length S plus S2 proteins for “cell”; the ratio of S2 to HIV-1 p24 for “supernatant”) ( D ) are shown. M, mock (empty vector-transfected). ( E ) Flow cytometry. The summarized results of the surface S expression are shown. ( F ) SARS-CoV-2 S-based fusion assay. The fusion activity was measured as described in STAR⍰METHODS, and fusion activity (arbitrary units) is shown. For the target cells, HEK293 cells expressing ACE2 and TMPRSS2 (filled) and HEK293 cells expressing ACE2 (open) were used. The results for Omicron S (top) or B.1 S (bottom) are shown in other panels as green and black lines, respectively. The results in HEK293-ACE2/TMPRSS2 cells and HEK293-ACE2 cells are shown as normal and broken lines, respectively. Data are expressed as the mean with SD. Assays were performed in quadruplicate ( B ) or triplicate ( D-F ). In B, D and E , each dot indicates the result of an individual replicate. Statistically significant differences (*P<0.05) versus the respective parental S [Omicron S (pseudovirus 2, top panels) or B.1 S (spike 1, bottom panels)] were determined by two-sided Student’s t test ( B and E ) or two-sided paired t test ( D ). In F , statistically significant differences (*FWERs<0.05) versus the respective parental S [Omicron S (spike 2, top panels) or B.1 S (spike 1, bottom panels)] through timepoints were determined by multiple regression. FWERs were calculated using the Holm method. See also .

    Techniques Used: Infection, Western Blot, Expressing, Plasmid Preparation, Transfection, Flow Cytometry, Single Vesicle Fusion Assay, Activity Assay

    mouse anti hiv 1 p24 monoclonal antibody  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc mouse anti hiv 1 p24 monoclonal antibody
    a , Western blotting of pseudoviruses. (Left) Representative blots of SARS-CoV-2 full-length S and cleaved S2 proteins as well as <t>HIV-1</t> <t>p24</t> capsid as an internal control. kDa, kilodaltons. (Right) The ratio of S2 to the full-length S plus S2 proteins on pseudovirus particles. Assays were performed in triplicate. Data are mean ± S.D. A statistically significant difference (*, P < 0.05) versus D614G S was determined by two-sided Student’s t test. b , Pseudovirus assay. The HIV-1-based reporter virus pseudotyped with SARS-CoV-2 S D614G or D614G/P681R was inoculated into HOS-ACE2 cells or HOS-ACE2/TMPRSS2 cells at 4 different doses (125, 250, 500 and 1,000 ng HIV-1 p24 antigen). Rates of infectivity compared to the virus pseudotyped with parental S D614G (1,000 ng HIV-1 p24) in HOS-ACE2 cells are shown. The labels above the HOS-ACE2/TMPRSS2 bars indicate the fold change versus the corresponding HOS-ACE2. Assays were performed in quadruplicate. c , Expression of S protein on the cell surface. (Left) Representative histogram of S protein expression on the cell surface. The number in the histogram indicates the mean fluorescence intensity (MFI). (Right) The MFI of surface S on the S-expressing cells. Assays were performed in triplicate. d , e , The kinetics of fusion velocity. d , Fitting of a mathematical model based on the kinetics of fusion activity data (see ). Each line indicates the result of respective mathematical model on the experimental data (shown in Fig. ). e , Initial velocity of the S-mediated fusion. Assays were performed in quadruplicate. Data are mean ± S.D. Statistically significant differences (* P < 0.05) were determined by two-sided, unpaired Student’s t test without adjustments for multiple comparisons ( b ), two-sided Student’s t test ( c ) or two-sided Welch’s t test ( e ). NS, no statistical significance.
    Mouse Anti Hiv 1 P24 Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti hiv 1 p24 monoclonal antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti hiv 1 p24 monoclonal antibody - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Enhanced fusogenicity and pathogenicity of SARS-CoV-2 Delta P681R mutation"

    Article Title: Enhanced fusogenicity and pathogenicity of SARS-CoV-2 Delta P681R mutation

    Journal: Nature

    doi: 10.1038/s41586-021-04266-9

    a , Western blotting of pseudoviruses. (Left) Representative blots of SARS-CoV-2 full-length S and cleaved S2 proteins as well as HIV-1 p24 capsid as an internal control. kDa, kilodaltons. (Right) The ratio of S2 to the full-length S plus S2 proteins on pseudovirus particles. Assays were performed in triplicate. Data are mean ± S.D. A statistically significant difference (*, P < 0.05) versus D614G S was determined by two-sided Student’s t test. b , Pseudovirus assay. The HIV-1-based reporter virus pseudotyped with SARS-CoV-2 S D614G or D614G/P681R was inoculated into HOS-ACE2 cells or HOS-ACE2/TMPRSS2 cells at 4 different doses (125, 250, 500 and 1,000 ng HIV-1 p24 antigen). Rates of infectivity compared to the virus pseudotyped with parental S D614G (1,000 ng HIV-1 p24) in HOS-ACE2 cells are shown. The labels above the HOS-ACE2/TMPRSS2 bars indicate the fold change versus the corresponding HOS-ACE2. Assays were performed in quadruplicate. c , Expression of S protein on the cell surface. (Left) Representative histogram of S protein expression on the cell surface. The number in the histogram indicates the mean fluorescence intensity (MFI). (Right) The MFI of surface S on the S-expressing cells. Assays were performed in triplicate. d , e , The kinetics of fusion velocity. d , Fitting of a mathematical model based on the kinetics of fusion activity data (see ). Each line indicates the result of respective mathematical model on the experimental data (shown in Fig. ). e , Initial velocity of the S-mediated fusion. Assays were performed in quadruplicate. Data are mean ± S.D. Statistically significant differences (* P < 0.05) were determined by two-sided, unpaired Student’s t test without adjustments for multiple comparisons ( b ), two-sided Student’s t test ( c ) or two-sided Welch’s t test ( e ). NS, no statistical significance.
    Figure Legend Snippet: a , Western blotting of pseudoviruses. (Left) Representative blots of SARS-CoV-2 full-length S and cleaved S2 proteins as well as HIV-1 p24 capsid as an internal control. kDa, kilodaltons. (Right) The ratio of S2 to the full-length S plus S2 proteins on pseudovirus particles. Assays were performed in triplicate. Data are mean ± S.D. A statistically significant difference (*, P < 0.05) versus D614G S was determined by two-sided Student’s t test. b , Pseudovirus assay. The HIV-1-based reporter virus pseudotyped with SARS-CoV-2 S D614G or D614G/P681R was inoculated into HOS-ACE2 cells or HOS-ACE2/TMPRSS2 cells at 4 different doses (125, 250, 500 and 1,000 ng HIV-1 p24 antigen). Rates of infectivity compared to the virus pseudotyped with parental S D614G (1,000 ng HIV-1 p24) in HOS-ACE2 cells are shown. The labels above the HOS-ACE2/TMPRSS2 bars indicate the fold change versus the corresponding HOS-ACE2. Assays were performed in quadruplicate. c , Expression of S protein on the cell surface. (Left) Representative histogram of S protein expression on the cell surface. The number in the histogram indicates the mean fluorescence intensity (MFI). (Right) The MFI of surface S on the S-expressing cells. Assays were performed in triplicate. d , e , The kinetics of fusion velocity. d , Fitting of a mathematical model based on the kinetics of fusion activity data (see ). Each line indicates the result of respective mathematical model on the experimental data (shown in Fig. ). e , Initial velocity of the S-mediated fusion. Assays were performed in quadruplicate. Data are mean ± S.D. Statistically significant differences (* P < 0.05) were determined by two-sided, unpaired Student’s t test without adjustments for multiple comparisons ( b ), two-sided Student’s t test ( c ) or two-sided Welch’s t test ( e ). NS, no statistical significance.

    Techniques Used: Western Blot, Infection, Expressing, Fluorescence, Activity Assay

    mouse anti hiv 1 p24 monoclonal antibody  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc mouse anti hiv 1 p24 monoclonal antibody
    a, Western blotting of pseudoviruses. (Left) Representative blots of SARS-CoV-2 full-length S and cleaved S2 proteins as well as <t>HIV-1</t> <t>p24</t> capsid as an internal control. kDa, kilodalton. (Right) The ratio of S2 to the full-length S plus S2 proteins on pseudovirus particles. b, Pseudovirus assay. The HIV-1-based reporter virus pseudotyped with the SARS-CoV-2 S D614G or D614G/P681R was inoculated into HOS-ACE2 cells or HOS-ACE2/TMPRSS2 cells at 4 different doses (125, 250, 500 and 1,000 ng HIV-1 p24 antigen). Percentages of infectivity compared to the virus pseudotyped with parental S D614G (1,000 ng HIV-1 p24) in HOS-ACE2 cells are shown. The numbers on the bars of the HOS-ACE2/TMPRSS2 cell data indicate the fold change versus the HOS-ACE2 cell data. Assays were performed in quadruplicate. NS, no statistical significance.
    Mouse Anti Hiv 1 P24 Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti hiv 1 p24 monoclonal antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti hiv 1 p24 monoclonal antibody - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "SARS-CoV-2 spike P681R mutation, a hallmark of the Delta variant, enhances viral fusogenicity and pathogenicity"

    Article Title: SARS-CoV-2 spike P681R mutation, a hallmark of the Delta variant, enhances viral fusogenicity and pathogenicity

    Journal: bioRxiv

    doi: 10.1101/2021.06.17.448820

    a, Western blotting of pseudoviruses. (Left) Representative blots of SARS-CoV-2 full-length S and cleaved S2 proteins as well as HIV-1 p24 capsid as an internal control. kDa, kilodalton. (Right) The ratio of S2 to the full-length S plus S2 proteins on pseudovirus particles. b, Pseudovirus assay. The HIV-1-based reporter virus pseudotyped with the SARS-CoV-2 S D614G or D614G/P681R was inoculated into HOS-ACE2 cells or HOS-ACE2/TMPRSS2 cells at 4 different doses (125, 250, 500 and 1,000 ng HIV-1 p24 antigen). Percentages of infectivity compared to the virus pseudotyped with parental S D614G (1,000 ng HIV-1 p24) in HOS-ACE2 cells are shown. The numbers on the bars of the HOS-ACE2/TMPRSS2 cell data indicate the fold change versus the HOS-ACE2 cell data. Assays were performed in quadruplicate. NS, no statistical significance.
    Figure Legend Snippet: a, Western blotting of pseudoviruses. (Left) Representative blots of SARS-CoV-2 full-length S and cleaved S2 proteins as well as HIV-1 p24 capsid as an internal control. kDa, kilodalton. (Right) The ratio of S2 to the full-length S plus S2 proteins on pseudovirus particles. b, Pseudovirus assay. The HIV-1-based reporter virus pseudotyped with the SARS-CoV-2 S D614G or D614G/P681R was inoculated into HOS-ACE2 cells or HOS-ACE2/TMPRSS2 cells at 4 different doses (125, 250, 500 and 1,000 ng HIV-1 p24 antigen). Percentages of infectivity compared to the virus pseudotyped with parental S D614G (1,000 ng HIV-1 p24) in HOS-ACE2 cells are shown. The numbers on the bars of the HOS-ACE2/TMPRSS2 cell data indicate the fold change versus the HOS-ACE2 cell data. Assays were performed in quadruplicate. NS, no statistical significance.

    Techniques Used: Western Blot, Infection

    anti phospho ikk2 ser180  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc anti phospho ikk2 ser180
    The effect of LOC645166 overexpressed in Jurkat cells on <t>IKK2</t> activation in response to TNF-α treatment. (A) The effect of LOC645166 on recruitment of the IKK complex to the K63-linked polyubiquitin chains. The co-immunoprecipitated proteins by anti-IKK2 monoclonal antibody were analyzed by western blotting, probed for K63-linked polyubiquitin chains by using anti-K63-linkage specific polyubiquitin monoclonal antibody or IKK2 by anti-IKK2 monoclonal antibody. (B) The effect of LOC645166 overexpression on the IKK2 phosphorylation. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector are set to 100%. (C) The effect of LOC645166 overexpression on the IKK2 phosphorylation in response to treatment with TNF-α. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector plus TNF-α treatment are set to 100%.
    Anti Phospho Ikk2 Ser180, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho ikk2 ser180/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho ikk2 ser180 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Down-Regulation of LOC645166 in T Cells of Ankylosing Spondylitis Patients Promotes the NF-κB Signaling via Decreasingly Blocking Recruitment of the IKK Complex to K63-Linked Polyubiquitin Chains"

    Article Title: Down-Regulation of LOC645166 in T Cells of Ankylosing Spondylitis Patients Promotes the NF-κB Signaling via Decreasingly Blocking Recruitment of the IKK Complex to K63-Linked Polyubiquitin Chains

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2021.591706

    The effect of LOC645166 overexpressed in Jurkat cells on IKK2 activation in response to TNF-α treatment. (A) The effect of LOC645166 on recruitment of the IKK complex to the K63-linked polyubiquitin chains. The co-immunoprecipitated proteins by anti-IKK2 monoclonal antibody were analyzed by western blotting, probed for K63-linked polyubiquitin chains by using anti-K63-linkage specific polyubiquitin monoclonal antibody or IKK2 by anti-IKK2 monoclonal antibody. (B) The effect of LOC645166 overexpression on the IKK2 phosphorylation. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector are set to 100%. (C) The effect of LOC645166 overexpression on the IKK2 phosphorylation in response to treatment with TNF-α. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector plus TNF-α treatment are set to 100%.
    Figure Legend Snippet: The effect of LOC645166 overexpressed in Jurkat cells on IKK2 activation in response to TNF-α treatment. (A) The effect of LOC645166 on recruitment of the IKK complex to the K63-linked polyubiquitin chains. The co-immunoprecipitated proteins by anti-IKK2 monoclonal antibody were analyzed by western blotting, probed for K63-linked polyubiquitin chains by using anti-K63-linkage specific polyubiquitin monoclonal antibody or IKK2 by anti-IKK2 monoclonal antibody. (B) The effect of LOC645166 overexpression on the IKK2 phosphorylation. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector are set to 100%. (C) The effect of LOC645166 overexpression on the IKK2 phosphorylation in response to treatment with TNF-α. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector plus TNF-α treatment are set to 100%.

    Techniques Used: Activation Assay, Immunoprecipitation, Western Blot, Over Expression, Transfection, Plasmid Preparation

    anti pikkα ser180  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc anti pikkα ser180
    Anti Pikkα Ser180, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti pikkα ser180/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti pikkα ser180 - by Bioz Stars, 2023-01
    94/100 stars

    Images

    phospho btk  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc phospho btk
    Effects of Rosae Multiflorae fructus extract (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-induced intracellular signaling. Bone marrow-derived macrophages (BMMs) were pretreated with/without RMF-E (20 µg/mL) for 2 hr in the presence of macrophage colony-stimulating factor (M-CSF) (30 ng/mL), and then RANKL (100 ng/mL)-treated, to stimulate intracellular signaling, at indicated times. Lysate (30 µg) was subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis and analyzed by immunoblotting. (A) The activation of mitogen-activated protein kinases (extracellular signal-regulated <t>kinase</t> <t>[ERK],</t> c-JUN N-terminal kinase [JNK], and p38) and IκBα was examined using their respective antibodies. (B) Twenty four-hr RANKL-stimulated BMMs were acutely treated with RMF-E (20 µg/mL). RANKL-stimulated Ca 2+ -oscillation by was measured using the fluorescence Ca 2+ indicator (Fura-2, AM). (C) Bruton's tyrosine kinase <t>(BTK)</t> and phospholipase C-γ2 (PLCγ2) activation were detected using the anti p-BTK/BTK and p-PLCγ2/PLCγ2 antibody, respectively. Fold change normalized by their non-phosphorylated proteins is presented in the right panel. Data are expressed as mean±standard deviation and are representative of 3 independent experiments. * P <0.05 and ** P <0.01 vs. the control group (0 µg/mL RMF-E).
    Phospho Btk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho btk/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho btk - by Bioz Stars, 2023-01
    94/100 stars

    Images

    1) Product Images from "Inhibitory Effect of Rosae Multiflorae Fructus Extracts on the Receptor Activator of NF-κB Ligand-Induced Osteoclastogenesis through Modulation of P38- and Ca 2+ -Mediated Nuclear Factor of Activated T-Cells Cytoplasmic 1 Expression"

    Article Title: Inhibitory Effect of Rosae Multiflorae Fructus Extracts on the Receptor Activator of NF-κB Ligand-Induced Osteoclastogenesis through Modulation of P38- and Ca 2+ -Mediated Nuclear Factor of Activated T-Cells Cytoplasmic 1 Expression

    Journal: Journal of Bone Metabolism

    doi: 10.11005/jbm.2020.27.1.53

    Effects of Rosae Multiflorae fructus extract (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-induced intracellular signaling. Bone marrow-derived macrophages (BMMs) were pretreated with/without RMF-E (20 µg/mL) for 2 hr in the presence of macrophage colony-stimulating factor (M-CSF) (30 ng/mL), and then RANKL (100 ng/mL)-treated, to stimulate intracellular signaling, at indicated times. Lysate (30 µg) was subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis and analyzed by immunoblotting. (A) The activation of mitogen-activated protein kinases (extracellular signal-regulated kinase [ERK], c-JUN N-terminal kinase [JNK], and p38) and IκBα was examined using their respective antibodies. (B) Twenty four-hr RANKL-stimulated BMMs were acutely treated with RMF-E (20 µg/mL). RANKL-stimulated Ca 2+ -oscillation by was measured using the fluorescence Ca 2+ indicator (Fura-2, AM). (C) Bruton's tyrosine kinase (BTK) and phospholipase C-γ2 (PLCγ2) activation were detected using the anti p-BTK/BTK and p-PLCγ2/PLCγ2 antibody, respectively. Fold change normalized by their non-phosphorylated proteins is presented in the right panel. Data are expressed as mean±standard deviation and are representative of 3 independent experiments. * P <0.05 and ** P <0.01 vs. the control group (0 µg/mL RMF-E).
    Figure Legend Snippet: Effects of Rosae Multiflorae fructus extract (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-induced intracellular signaling. Bone marrow-derived macrophages (BMMs) were pretreated with/without RMF-E (20 µg/mL) for 2 hr in the presence of macrophage colony-stimulating factor (M-CSF) (30 ng/mL), and then RANKL (100 ng/mL)-treated, to stimulate intracellular signaling, at indicated times. Lysate (30 µg) was subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis and analyzed by immunoblotting. (A) The activation of mitogen-activated protein kinases (extracellular signal-regulated kinase [ERK], c-JUN N-terminal kinase [JNK], and p38) and IκBα was examined using their respective antibodies. (B) Twenty four-hr RANKL-stimulated BMMs were acutely treated with RMF-E (20 µg/mL). RANKL-stimulated Ca 2+ -oscillation by was measured using the fluorescence Ca 2+ indicator (Fura-2, AM). (C) Bruton's tyrosine kinase (BTK) and phospholipase C-γ2 (PLCγ2) activation were detected using the anti p-BTK/BTK and p-PLCγ2/PLCγ2 antibody, respectively. Fold change normalized by their non-phosphorylated proteins is presented in the right panel. Data are expressed as mean±standard deviation and are representative of 3 independent experiments. * P <0.05 and ** P <0.01 vs. the control group (0 µg/mL RMF-E).

    Techniques Used: Derivative Assay, Polyacrylamide Gel Electrophoresis, Western Blot, Activation Assay, Fluorescence, Standard Deviation

    Schematic diagram of the effect of Rosae Multiflorae fructus extract (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis. The RANKL/RANK interaction may lead to mitogen-activated protein kinases (MAPKs) activation, followed by induction of c-Fos expression, and it also leads to Bruton's tyrosine kinase (BTK) and phospholipase C-γ2 (PLCγ2) activation inducing calcium signaling, followed by induction of nuclear factor-activated T cells c1 (NFATc1) expression and activation. RMF-E may inhibit NFATc1 induction via suppression of c-Fos and modulation of the BTK/PLCγ signaling pathways regulating Ca 2+ signaling, resulting in the inhibition of RANKL-induced osteoclastogenesis. The red line indicates the inhibition pathway of RMF-E. TRAF6, tumor necrosis factor receptor associated factor 6; ITAM, immunoreceptor tyrosine-based activation motif.
    Figure Legend Snippet: Schematic diagram of the effect of Rosae Multiflorae fructus extract (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis. The RANKL/RANK interaction may lead to mitogen-activated protein kinases (MAPKs) activation, followed by induction of c-Fos expression, and it also leads to Bruton's tyrosine kinase (BTK) and phospholipase C-γ2 (PLCγ2) activation inducing calcium signaling, followed by induction of nuclear factor-activated T cells c1 (NFATc1) expression and activation. RMF-E may inhibit NFATc1 induction via suppression of c-Fos and modulation of the BTK/PLCγ signaling pathways regulating Ca 2+ signaling, resulting in the inhibition of RANKL-induced osteoclastogenesis. The red line indicates the inhibition pathway of RMF-E. TRAF6, tumor necrosis factor receptor associated factor 6; ITAM, immunoreceptor tyrosine-based activation motif.

    Techniques Used: Activation Assay, Expressing, Inhibition

    phospho  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc phospho
    Phospho, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho - by Bioz Stars, 2023-01
    94/100 stars

    Images

    phospho  (Cell Signaling Technology Inc)


    Bioz Verified Symbol Cell Signaling Technology Inc is a verified supplier
    Bioz Manufacturer Symbol Cell Signaling Technology Inc manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94

    Structured Review

    Cell Signaling Technology Inc phospho
    Phospho, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho - by Bioz Stars, 2023-01
    94/100 stars

    Images

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Cell Signaling Technology Inc phospho foxo
    17-AAG induces downregulation of <t>FOXO</t> and ERK1/2 signaling mediators in bladder cancer cells. Detection of total and constitutively phosphorylated protein levels of FOXO transcription factors and <t>Erk1/2</t> <t>(p44/42)</t> kinases in all three bladder cancer cell lines, upon 17-AAG administration for 24 hours, via a Western blotting approach. All immunoblotting experiments were repeated three times.
    Phospho Foxo, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho foxo/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho foxo - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc mouse anti hiv 1 p24 monoclonal antibody
    Virological features of BA.2 in vitro (A) Scheme for the chimeric recombinant SARS-CoV-2 used in this study. The SARS-CoV-2 genome and its genes are shown. The template was SARS-CoV-2 strain WK-521 (PANGO lineage A, GISAID ID: EPI_ISL_408667), and the S genes were swapped with those of the respective lineages/strains (GISAID IDs are indicated in the figure). ORF7a was swapped with the sfGFP gene. (B) Growth kinetics of chimeric recombinant SARS-CoV-2 in Vero cells, VeroE6/TMPRSS2 cells, Calu-3 cells, and human nasal epithelial cells. (C) Fluorescence microscopy. The GFP area was measured in infected VeroE6/TMPRSS2 cells (multiplicity of infection [m.o.i.] 0.01) at 48 h.p.i. Left, representative panels. Higher-magnification views of the regions indicated by squares are shown at the bottom. Representative time-course data are shown in <xref ref-type=Figure S2 E. Scale bars, 500 μm. Right, the summarized results. The numbers in the panel indicate the numbers of GFP-positive cells counted. (D) Plaque assay. Representative panels (left) and a summary of the recorded plaque diameters (20 plaques per virus) (right) are shown. (E) S expression on the cell surface. Representative histograms stained with an anti-S1/S2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates mean fluorescence intensity (MFI). Gray histograms indicate isotype controls. (F) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (G) Western blotting. Left, representative blots of S-expressing cells (top) and pseudovirus (bottom). ACTB is an internal control for the cells, whereas HIV-1 p24 is an internal control for the pseudovirus. kDa, kilodalton. Middle, the ratio of S2 to the full-length S plus S2 proteins in the cells. Right, the ratio of S2 to HIV-1 p24 in the pseudovirus (supernatant). (H) Pseudovirus assay. The percent infectivity compared with that of the virus pseudotyped with B.1.1 S are shown. (I) Binding affinity of SARS-CoV-2 S RBD to ACE2 by yeast surface display. The percentage of SARS-CoV-2 S RBD expressed on yeast binding to soluble ACE2 (left) and the summarized K D values (right) are shown. (J) TMPRSS2 expression on the cell surface. Representative histograms stained with an anti-TMPRSS2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates MFI. Gray histograms indicate the isotype controls. (K) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (L) Fold increase in pseudovirus infectivity based on TMPRSS2 expression. (M) E64d treatment. IC50, 50% inhibitory concentration; ND, not determined. (N) Growth kinetics of chimeric recombinant SARS-CoV-2 in HK293-ACE2 and HEK293-ACE2/TMPRSS2 cells. Assays were performed in quadruplicate (B, H, L, J, and N), octuplicate (B, most right) or triplicate (E–G, I, J, K, and M), and the presented data are expressed as the average ± SD. Each dot indicates the result of an individual plaque (D) and an individual replicate (E, G– J, L and I). Statistically significant differences between BA.2 and other variants across time points were determined by multiple regression (B, F, K, and N). Family-wise error rates (FWERs) calculated using the Holm method are indicated in the figures. Statistically significant differences between BA.1 and BA.2 were determined by two-sided Mann-Whitney U tests (C and D), two-sided Student’s t tests (E, H, and I), or two-sided paired t test (G). See also Figure S3 . " width="250" height="auto" />
    Mouse Anti Hiv 1 P24 Monoclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mouse anti hiv 1 p24 monoclonal antibody/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mouse anti hiv 1 p24 monoclonal antibody - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc anti phospho ikk2 ser180
    The effect of LOC645166 overexpressed in Jurkat cells on <t>IKK2</t> activation in response to TNF-α treatment. (A) The effect of LOC645166 on recruitment of the IKK complex to the K63-linked polyubiquitin chains. The co-immunoprecipitated proteins by anti-IKK2 monoclonal antibody were analyzed by western blotting, probed for K63-linked polyubiquitin chains by using anti-K63-linkage specific polyubiquitin monoclonal antibody or IKK2 by anti-IKK2 monoclonal antibody. (B) The effect of LOC645166 overexpression on the IKK2 phosphorylation. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector are set to 100%. (C) The effect of LOC645166 overexpression on the IKK2 phosphorylation in response to treatment with TNF-α. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector plus TNF-α treatment are set to 100%.
    Anti Phospho Ikk2 Ser180, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti phospho ikk2 ser180/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti phospho ikk2 ser180 - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc anti pikkα ser180
    The effect of LOC645166 overexpressed in Jurkat cells on <t>IKK2</t> activation in response to TNF-α treatment. (A) The effect of LOC645166 on recruitment of the IKK complex to the K63-linked polyubiquitin chains. The co-immunoprecipitated proteins by anti-IKK2 monoclonal antibody were analyzed by western blotting, probed for K63-linked polyubiquitin chains by using anti-K63-linkage specific polyubiquitin monoclonal antibody or IKK2 by anti-IKK2 monoclonal antibody. (B) The effect of LOC645166 overexpression on the IKK2 phosphorylation. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector are set to 100%. (C) The effect of LOC645166 overexpression on the IKK2 phosphorylation in response to treatment with TNF-α. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector plus TNF-α treatment are set to 100%.
    Anti Pikkα Ser180, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/anti pikkα ser180/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    anti pikkα ser180 - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc phospho btk
    Effects of Rosae Multiflorae fructus extract (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-induced intracellular signaling. Bone marrow-derived macrophages (BMMs) were pretreated with/without RMF-E (20 µg/mL) for 2 hr in the presence of macrophage colony-stimulating factor (M-CSF) (30 ng/mL), and then RANKL (100 ng/mL)-treated, to stimulate intracellular signaling, at indicated times. Lysate (30 µg) was subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis and analyzed by immunoblotting. (A) The activation of mitogen-activated protein kinases (extracellular signal-regulated <t>kinase</t> <t>[ERK],</t> c-JUN N-terminal kinase [JNK], and p38) and IκBα was examined using their respective antibodies. (B) Twenty four-hr RANKL-stimulated BMMs were acutely treated with RMF-E (20 µg/mL). RANKL-stimulated Ca 2+ -oscillation by was measured using the fluorescence Ca 2+ indicator (Fura-2, AM). (C) Bruton's tyrosine kinase <t>(BTK)</t> and phospholipase C-γ2 (PLCγ2) activation were detected using the anti p-BTK/BTK and p-PLCγ2/PLCγ2 antibody, respectively. Fold change normalized by their non-phosphorylated proteins is presented in the right panel. Data are expressed as mean±standard deviation and are representative of 3 independent experiments. * P <0.05 and ** P <0.01 vs. the control group (0 µg/mL RMF-E).
    Phospho Btk, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho btk/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho btk - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    94
    Cell Signaling Technology Inc phospho
    Effects of Rosae Multiflorae fructus extract (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-induced intracellular signaling. Bone marrow-derived macrophages (BMMs) were pretreated with/without RMF-E (20 µg/mL) for 2 hr in the presence of macrophage colony-stimulating factor (M-CSF) (30 ng/mL), and then RANKL (100 ng/mL)-treated, to stimulate intracellular signaling, at indicated times. Lysate (30 µg) was subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis and analyzed by immunoblotting. (A) The activation of mitogen-activated protein kinases (extracellular signal-regulated <t>kinase</t> <t>[ERK],</t> c-JUN N-terminal kinase [JNK], and p38) and IκBα was examined using their respective antibodies. (B) Twenty four-hr RANKL-stimulated BMMs were acutely treated with RMF-E (20 µg/mL). RANKL-stimulated Ca 2+ -oscillation by was measured using the fluorescence Ca 2+ indicator (Fura-2, AM). (C) Bruton's tyrosine kinase <t>(BTK)</t> and phospholipase C-γ2 (PLCγ2) activation were detected using the anti p-BTK/BTK and p-PLCγ2/PLCγ2 antibody, respectively. Fold change normalized by their non-phosphorylated proteins is presented in the right panel. Data are expressed as mean±standard deviation and are representative of 3 independent experiments. * P <0.05 and ** P <0.01 vs. the control group (0 µg/mL RMF-E).
    Phospho, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/phospho/product/Cell Signaling Technology Inc
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    phospho - by Bioz Stars, 2023-01
    94/100 stars
      Buy from Supplier

    Image Search Results


    17-AAG induces downregulation of FOXO and ERK1/2 signaling mediators in bladder cancer cells. Detection of total and constitutively phosphorylated protein levels of FOXO transcription factors and Erk1/2 (p44/42) kinases in all three bladder cancer cell lines, upon 17-AAG administration for 24 hours, via a Western blotting approach. All immunoblotting experiments were repeated three times.

    Journal: BMC Cancer

    Article Title: 17-Allylamino-17-demethoxygeldanamycin induces downregulation of critical Hsp90 protein clients and results in cell cycle arrest and apoptosis of human urinary bladder cancer cells

    doi: 10.1186/1471-2407-10-481

    Figure Lengend Snippet: 17-AAG induces downregulation of FOXO and ERK1/2 signaling mediators in bladder cancer cells. Detection of total and constitutively phosphorylated protein levels of FOXO transcription factors and Erk1/2 (p44/42) kinases in all three bladder cancer cell lines, upon 17-AAG administration for 24 hours, via a Western blotting approach. All immunoblotting experiments were repeated three times.

    Article Snippet: Polyclonal and monoclonal antibodies against Caspase-8, Caspase-9, Caspase-3, PARP, Lamin A/C, phospho-Akt (Ser473), phospho-Akt (Thr308), Akt, phospho-IGF-ΙRβ (Tyr1131), IGF-ΙRα, FOXO, phospho-FOXO, phospho-IKKα/β (Ser180/Ser181), IKKα, IKKβ, phospho-p44/42 (Thr202/Tyr204), p44/42, α-tubulin, phospho-c-Met (Tyr1234/Tyr1235), c-Met, CHIP and pan-actin were purchased from Cell Signaling Technology Inc. (Hertfordshire, UK), whereas antibodies against Hsp90α/β, Hsp70, Cdk4, pRb, E2F1 and NF-κB (p65) were supplied by Santa Cruz Biotechnology Inc. (California, USA).

    Techniques: Western Blot

    Virological features of BA.2 in vitro (A) Scheme for the chimeric recombinant SARS-CoV-2 used in this study. The SARS-CoV-2 genome and its genes are shown. The template was SARS-CoV-2 strain WK-521 (PANGO lineage A, GISAID ID: EPI_ISL_408667), and the S genes were swapped with those of the respective lineages/strains (GISAID IDs are indicated in the figure). ORF7a was swapped with the sfGFP gene. (B) Growth kinetics of chimeric recombinant SARS-CoV-2 in Vero cells, VeroE6/TMPRSS2 cells, Calu-3 cells, and human nasal epithelial cells. (C) Fluorescence microscopy. The GFP area was measured in infected VeroE6/TMPRSS2 cells (multiplicity of infection [m.o.i.] 0.01) at 48 h.p.i. Left, representative panels. Higher-magnification views of the regions indicated by squares are shown at the bottom. Representative time-course data are shown in <xref ref-type=Figure S2 E. Scale bars, 500 μm. Right, the summarized results. The numbers in the panel indicate the numbers of GFP-positive cells counted. (D) Plaque assay. Representative panels (left) and a summary of the recorded plaque diameters (20 plaques per virus) (right) are shown. (E) S expression on the cell surface. Representative histograms stained with an anti-S1/S2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates mean fluorescence intensity (MFI). Gray histograms indicate isotype controls. (F) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (G) Western blotting. Left, representative blots of S-expressing cells (top) and pseudovirus (bottom). ACTB is an internal control for the cells, whereas HIV-1 p24 is an internal control for the pseudovirus. kDa, kilodalton. Middle, the ratio of S2 to the full-length S plus S2 proteins in the cells. Right, the ratio of S2 to HIV-1 p24 in the pseudovirus (supernatant). (H) Pseudovirus assay. The percent infectivity compared with that of the virus pseudotyped with B.1.1 S are shown. (I) Binding affinity of SARS-CoV-2 S RBD to ACE2 by yeast surface display. The percentage of SARS-CoV-2 S RBD expressed on yeast binding to soluble ACE2 (left) and the summarized K D values (right) are shown. (J) TMPRSS2 expression on the cell surface. Representative histograms stained with an anti-TMPRSS2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates MFI. Gray histograms indicate the isotype controls. (K) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (L) Fold increase in pseudovirus infectivity based on TMPRSS2 expression. (M) E64d treatment. IC50, 50% inhibitory concentration; ND, not determined. (N) Growth kinetics of chimeric recombinant SARS-CoV-2 in HK293-ACE2 and HEK293-ACE2/TMPRSS2 cells. Assays were performed in quadruplicate (B, H, L, J, and N), octuplicate (B, most right) or triplicate (E–G, I, J, K, and M), and the presented data are expressed as the average ± SD. Each dot indicates the result of an individual plaque (D) and an individual replicate (E, G– J, L and I). Statistically significant differences between BA.2 and other variants across time points were determined by multiple regression (B, F, K, and N). Family-wise error rates (FWERs) calculated using the Holm method are indicated in the figures. Statistically significant differences between BA.1 and BA.2 were determined by two-sided Mann-Whitney U tests (C and D), two-sided Student’s t tests (E, H, and I), or two-sided paired t test (G). See also Figure S3 . " width="100%" height="100%">

    Journal: Cell

    Article Title: Virological characteristics of the SARS-CoV-2 Omicron BA.2 spike

    doi: 10.1016/j.cell.2022.04.035

    Figure Lengend Snippet: Virological features of BA.2 in vitro (A) Scheme for the chimeric recombinant SARS-CoV-2 used in this study. The SARS-CoV-2 genome and its genes are shown. The template was SARS-CoV-2 strain WK-521 (PANGO lineage A, GISAID ID: EPI_ISL_408667), and the S genes were swapped with those of the respective lineages/strains (GISAID IDs are indicated in the figure). ORF7a was swapped with the sfGFP gene. (B) Growth kinetics of chimeric recombinant SARS-CoV-2 in Vero cells, VeroE6/TMPRSS2 cells, Calu-3 cells, and human nasal epithelial cells. (C) Fluorescence microscopy. The GFP area was measured in infected VeroE6/TMPRSS2 cells (multiplicity of infection [m.o.i.] 0.01) at 48 h.p.i. Left, representative panels. Higher-magnification views of the regions indicated by squares are shown at the bottom. Representative time-course data are shown in Figure S2 E. Scale bars, 500 μm. Right, the summarized results. The numbers in the panel indicate the numbers of GFP-positive cells counted. (D) Plaque assay. Representative panels (left) and a summary of the recorded plaque diameters (20 plaques per virus) (right) are shown. (E) S expression on the cell surface. Representative histograms stained with an anti-S1/S2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates mean fluorescence intensity (MFI). Gray histograms indicate isotype controls. (F) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (G) Western blotting. Left, representative blots of S-expressing cells (top) and pseudovirus (bottom). ACTB is an internal control for the cells, whereas HIV-1 p24 is an internal control for the pseudovirus. kDa, kilodalton. Middle, the ratio of S2 to the full-length S plus S2 proteins in the cells. Right, the ratio of S2 to HIV-1 p24 in the pseudovirus (supernatant). (H) Pseudovirus assay. The percent infectivity compared with that of the virus pseudotyped with B.1.1 S are shown. (I) Binding affinity of SARS-CoV-2 S RBD to ACE2 by yeast surface display. The percentage of SARS-CoV-2 S RBD expressed on yeast binding to soluble ACE2 (left) and the summarized K D values (right) are shown. (J) TMPRSS2 expression on the cell surface. Representative histograms stained with an anti-TMPRSS2 polyclonal antibody (left) and the summarized data (right) are shown. In the left panel, the number in the histogram indicates MFI. Gray histograms indicate the isotype controls. (K) S-based fusion assay. The recorded fusion activity (arbitrary units) is shown. (L) Fold increase in pseudovirus infectivity based on TMPRSS2 expression. (M) E64d treatment. IC50, 50% inhibitory concentration; ND, not determined. (N) Growth kinetics of chimeric recombinant SARS-CoV-2 in HK293-ACE2 and HEK293-ACE2/TMPRSS2 cells. Assays were performed in quadruplicate (B, H, L, J, and N), octuplicate (B, most right) or triplicate (E–G, I, J, K, and M), and the presented data are expressed as the average ± SD. Each dot indicates the result of an individual plaque (D) and an individual replicate (E, G– J, L and I). Statistically significant differences between BA.2 and other variants across time points were determined by multiple regression (B, F, K, and N). Family-wise error rates (FWERs) calculated using the Holm method are indicated in the figures. Statistically significant differences between BA.1 and BA.2 were determined by two-sided Mann-Whitney U tests (C and D), two-sided Student’s t tests (E, H, and I), or two-sided paired t test (G). See also Figure S3 .

    Article Snippet: The pelleted virions were resuspended in 1× NuPAGE LDS sample buffer (Thermo Fisher Scientific, Cat# NP0007) containing 2% β-mercaptoethanol and incubated at 70°C for 10 m. For protein detection, the following antibodies were used: mouse anti-SARS-CoV-2 S monoclonal antibody (clone 1A9, GeneTex, Cat# GTX632604, 1:10,000), mouse anti-HIV-1 p24 monoclonal antibody (clone 183-H12-5C, obtained from the Nih HIV Reagent Program, Cat# ARP-3537, 1:2,000), rabbit anti-beta actin (ACTB) monoclonal antibody (clone 13E5, Cell Signaling, Cat# 4970, 1:5,000), horseradish peroxidase (HRP)-conjugated donkey anti-rabbit IgG polyclonal antibody (Jackson ImmunoResearch, Cat# 711-035-152, 1:10,000) and HRP-conjugated donkey anti-mouse IgG polyclonal antibody (Jackson ImmunoResearch, Cat# 715-035-150, 1:10,000).

    Techniques: In Vitro, Recombinant, Fluorescence, Microscopy, Infection, Plaque Assay, Expressing, Staining, Single Vesicle Fusion Assay, Activity Assay, Western Blot, Binding Assay, Concentration Assay, MANN-WHITNEY

    Journal: Cell

    Article Title: Virological characteristics of the SARS-CoV-2 Omicron BA.2 spike

    doi: 10.1016/j.cell.2022.04.035

    Figure Lengend Snippet:

    Article Snippet: The pelleted virions were resuspended in 1× NuPAGE LDS sample buffer (Thermo Fisher Scientific, Cat# NP0007) containing 2% β-mercaptoethanol and incubated at 70°C for 10 m. For protein detection, the following antibodies were used: mouse anti-SARS-CoV-2 S monoclonal antibody (clone 1A9, GeneTex, Cat# GTX632604, 1:10,000), mouse anti-HIV-1 p24 monoclonal antibody (clone 183-H12-5C, obtained from the Nih HIV Reagent Program, Cat# ARP-3537, 1:2,000), rabbit anti-beta actin (ACTB) monoclonal antibody (clone 13E5, Cell Signaling, Cat# 4970, 1:5,000), horseradish peroxidase (HRP)-conjugated donkey anti-rabbit IgG polyclonal antibody (Jackson ImmunoResearch, Cat# 711-035-152, 1:10,000) and HRP-conjugated donkey anti-mouse IgG polyclonal antibody (Jackson ImmunoResearch, Cat# 715-035-150, 1:10,000).

    Techniques: Recombinant, Expressing, Protease Inhibitor, Activity Assay, Luciferase, CCK-8 Assay, Sequencing, Plasmid Preparation, Software

    The effect of LOC645166 overexpressed in Jurkat cells on IKK2 activation in response to TNF-α treatment. (A) The effect of LOC645166 on recruitment of the IKK complex to the K63-linked polyubiquitin chains. The co-immunoprecipitated proteins by anti-IKK2 monoclonal antibody were analyzed by western blotting, probed for K63-linked polyubiquitin chains by using anti-K63-linkage specific polyubiquitin monoclonal antibody or IKK2 by anti-IKK2 monoclonal antibody. (B) The effect of LOC645166 overexpression on the IKK2 phosphorylation. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector are set to 100%. (C) The effect of LOC645166 overexpression on the IKK2 phosphorylation in response to treatment with TNF-α. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector plus TNF-α treatment are set to 100%.

    Journal: Frontiers in Immunology

    Article Title: Down-Regulation of LOC645166 in T Cells of Ankylosing Spondylitis Patients Promotes the NF-κB Signaling via Decreasingly Blocking Recruitment of the IKK Complex to K63-Linked Polyubiquitin Chains

    doi: 10.3389/fimmu.2021.591706

    Figure Lengend Snippet: The effect of LOC645166 overexpressed in Jurkat cells on IKK2 activation in response to TNF-α treatment. (A) The effect of LOC645166 on recruitment of the IKK complex to the K63-linked polyubiquitin chains. The co-immunoprecipitated proteins by anti-IKK2 monoclonal antibody were analyzed by western blotting, probed for K63-linked polyubiquitin chains by using anti-K63-linkage specific polyubiquitin monoclonal antibody or IKK2 by anti-IKK2 monoclonal antibody. (B) The effect of LOC645166 overexpression on the IKK2 phosphorylation. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector are set to 100%. (C) The effect of LOC645166 overexpression on the IKK2 phosphorylation in response to treatment with TNF-α. The ratios of p-IKK2/actin averaged from four independent experiments are plotted. The ratios of p-IKK2/actin obtained from cells transfected with control vector plus TNF-α treatment are set to 100%.

    Article Snippet: Anti-IKK2, anti-phospho-IKK2 (Ser180), anti-STAT3, anti-phospho-STAT3 (Tyr705), anti-JAK2, anti-phospho-JAK2 (Tyr1007/1008), anti-ubiquitin, anti-K63-linkage specific polyubiquitin, anti-lamin A/C, anti-actin, anti-p50, anti-IκBα and anti-phospho-IκBα (Ser32) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Activation Assay, Immunoprecipitation, Western Blot, Over Expression, Transfection, Plasmid Preparation

    Effects of Rosae Multiflorae fructus extract (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-induced intracellular signaling. Bone marrow-derived macrophages (BMMs) were pretreated with/without RMF-E (20 µg/mL) for 2 hr in the presence of macrophage colony-stimulating factor (M-CSF) (30 ng/mL), and then RANKL (100 ng/mL)-treated, to stimulate intracellular signaling, at indicated times. Lysate (30 µg) was subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis and analyzed by immunoblotting. (A) The activation of mitogen-activated protein kinases (extracellular signal-regulated kinase [ERK], c-JUN N-terminal kinase [JNK], and p38) and IκBα was examined using their respective antibodies. (B) Twenty four-hr RANKL-stimulated BMMs were acutely treated with RMF-E (20 µg/mL). RANKL-stimulated Ca 2+ -oscillation by was measured using the fluorescence Ca 2+ indicator (Fura-2, AM). (C) Bruton's tyrosine kinase (BTK) and phospholipase C-γ2 (PLCγ2) activation were detected using the anti p-BTK/BTK and p-PLCγ2/PLCγ2 antibody, respectively. Fold change normalized by their non-phosphorylated proteins is presented in the right panel. Data are expressed as mean±standard deviation and are representative of 3 independent experiments. * P <0.05 and ** P <0.01 vs. the control group (0 µg/mL RMF-E).

    Journal: Journal of Bone Metabolism

    Article Title: Inhibitory Effect of Rosae Multiflorae Fructus Extracts on the Receptor Activator of NF-κB Ligand-Induced Osteoclastogenesis through Modulation of P38- and Ca 2+ -Mediated Nuclear Factor of Activated T-Cells Cytoplasmic 1 Expression

    doi: 10.11005/jbm.2020.27.1.53

    Figure Lengend Snippet: Effects of Rosae Multiflorae fructus extract (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-induced intracellular signaling. Bone marrow-derived macrophages (BMMs) were pretreated with/without RMF-E (20 µg/mL) for 2 hr in the presence of macrophage colony-stimulating factor (M-CSF) (30 ng/mL), and then RANKL (100 ng/mL)-treated, to stimulate intracellular signaling, at indicated times. Lysate (30 µg) was subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis and analyzed by immunoblotting. (A) The activation of mitogen-activated protein kinases (extracellular signal-regulated kinase [ERK], c-JUN N-terminal kinase [JNK], and p38) and IκBα was examined using their respective antibodies. (B) Twenty four-hr RANKL-stimulated BMMs were acutely treated with RMF-E (20 µg/mL). RANKL-stimulated Ca 2+ -oscillation by was measured using the fluorescence Ca 2+ indicator (Fura-2, AM). (C) Bruton's tyrosine kinase (BTK) and phospholipase C-γ2 (PLCγ2) activation were detected using the anti p-BTK/BTK and p-PLCγ2/PLCγ2 antibody, respectively. Fold change normalized by their non-phosphorylated proteins is presented in the right panel. Data are expressed as mean±standard deviation and are representative of 3 independent experiments. * P <0.05 and ** P <0.01 vs. the control group (0 µg/mL RMF-E).

    Article Snippet: Antibodies against other proteins used in this study (phospho-ERK [p-ERK; Thr202/Tyr204], ERK, phospho-JNK [Thr183/Tyr185], JNK, p-p38 [Thr180/Tyr182], p-38, phospho-IκBα [Ser32], IκBα, phospho-BTK [Ser180], BTK, phospho-PLCγ2 [Tyr759], and PLCγ2) were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Derivative Assay, Polyacrylamide Gel Electrophoresis, Western Blot, Activation Assay, Fluorescence, Standard Deviation

    Schematic diagram of the effect of Rosae Multiflorae fructus extract (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis. The RANKL/RANK interaction may lead to mitogen-activated protein kinases (MAPKs) activation, followed by induction of c-Fos expression, and it also leads to Bruton's tyrosine kinase (BTK) and phospholipase C-γ2 (PLCγ2) activation inducing calcium signaling, followed by induction of nuclear factor-activated T cells c1 (NFATc1) expression and activation. RMF-E may inhibit NFATc1 induction via suppression of c-Fos and modulation of the BTK/PLCγ signaling pathways regulating Ca 2+ signaling, resulting in the inhibition of RANKL-induced osteoclastogenesis. The red line indicates the inhibition pathway of RMF-E. TRAF6, tumor necrosis factor receptor associated factor 6; ITAM, immunoreceptor tyrosine-based activation motif.

    Journal: Journal of Bone Metabolism

    Article Title: Inhibitory Effect of Rosae Multiflorae Fructus Extracts on the Receptor Activator of NF-κB Ligand-Induced Osteoclastogenesis through Modulation of P38- and Ca 2+ -Mediated Nuclear Factor of Activated T-Cells Cytoplasmic 1 Expression

    doi: 10.11005/jbm.2020.27.1.53

    Figure Lengend Snippet: Schematic diagram of the effect of Rosae Multiflorae fructus extract (RMF-E) on receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis. The RANKL/RANK interaction may lead to mitogen-activated protein kinases (MAPKs) activation, followed by induction of c-Fos expression, and it also leads to Bruton's tyrosine kinase (BTK) and phospholipase C-γ2 (PLCγ2) activation inducing calcium signaling, followed by induction of nuclear factor-activated T cells c1 (NFATc1) expression and activation. RMF-E may inhibit NFATc1 induction via suppression of c-Fos and modulation of the BTK/PLCγ signaling pathways regulating Ca 2+ signaling, resulting in the inhibition of RANKL-induced osteoclastogenesis. The red line indicates the inhibition pathway of RMF-E. TRAF6, tumor necrosis factor receptor associated factor 6; ITAM, immunoreceptor tyrosine-based activation motif.

    Article Snippet: Antibodies against other proteins used in this study (phospho-ERK [p-ERK; Thr202/Tyr204], ERK, phospho-JNK [Thr183/Tyr185], JNK, p-p38 [Thr180/Tyr182], p-38, phospho-IκBα [Ser32], IκBα, phospho-BTK [Ser180], BTK, phospho-PLCγ2 [Tyr759], and PLCγ2) were purchased from Cell Signaling Technology (Danvers, MA, USA).

    Techniques: Activation Assay, Expressing, Inhibition