fitc  (Vector Laboratories)


Bioz Verified Symbol Vector Laboratories is a verified supplier
Bioz Manufacturer Symbol Vector Laboratories manufactures this product  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 86

    Structured Review

    Vector Laboratories fitc
    M cells are the major sites at which L-PTC breaches the intestinal epithelial barrier. ( a , b ) Mice were treated intraperitoneally with anti-RANKL antibody or control rat IgG. ( a ) Alexa Fluor 568-labelled L-PTC (red) was injected into ligated mouse intestinal loops and incubated for 2 h, and FAE regions were stained with <t>FITC-labelled</t> <t>UEA-1</t> (green) and iFluor 405-labelled phalloidin (blue). Scale bar, 20 μm. ( b ) L-PTC was intragastrically (1.0 pmol: 0.75 μg) or intraperitoneally (0.067 fmol: 50 pg) administered to mice treated with anti-RANKL antibody or rat IgG (intragastrically, n =13 per group; intraperitoneally, n =5 per group). Statistical analyses were performed with the log-rank test. NS, not significant. The data ( a ) are representative of two independent experiments.
    Fitc, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 86/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fitc/product/Vector Laboratories
    Average 86 stars, based on 40 article reviews
    Price from $9.99 to $1999.99
    fitc - by Bioz Stars, 2022-10
    86/100 stars

    Images

    1) Product Images from "Botulinum toxin A complex exploits intestinal M cells to enter the host and exert neurotoxicity"

    Article Title: Botulinum toxin A complex exploits intestinal M cells to enter the host and exert neurotoxicity

    Journal: Nature Communications

    doi: 10.1038/ncomms7255

    M cells are the major sites at which L-PTC breaches the intestinal epithelial barrier. ( a , b ) Mice were treated intraperitoneally with anti-RANKL antibody or control rat IgG. ( a ) Alexa Fluor 568-labelled L-PTC (red) was injected into ligated mouse intestinal loops and incubated for 2 h, and FAE regions were stained with FITC-labelled UEA-1 (green) and iFluor 405-labelled phalloidin (blue). Scale bar, 20 μm. ( b ) L-PTC was intragastrically (1.0 pmol: 0.75 μg) or intraperitoneally (0.067 fmol: 50 pg) administered to mice treated with anti-RANKL antibody or rat IgG (intragastrically, n =13 per group; intraperitoneally, n =5 per group). Statistical analyses were performed with the log-rank test. NS, not significant. The data ( a ) are representative of two independent experiments.
    Figure Legend Snippet: M cells are the major sites at which L-PTC breaches the intestinal epithelial barrier. ( a , b ) Mice were treated intraperitoneally with anti-RANKL antibody or control rat IgG. ( a ) Alexa Fluor 568-labelled L-PTC (red) was injected into ligated mouse intestinal loops and incubated for 2 h, and FAE regions were stained with FITC-labelled UEA-1 (green) and iFluor 405-labelled phalloidin (blue). Scale bar, 20 μm. ( b ) L-PTC was intragastrically (1.0 pmol: 0.75 μg) or intraperitoneally (0.067 fmol: 50 pg) administered to mice treated with anti-RANKL antibody or rat IgG (intragastrically, n =13 per group; intraperitoneally, n =5 per group). Statistical analyses were performed with the log-rank test. NS, not significant. The data ( a ) are representative of two independent experiments.

    Techniques Used: Mouse Assay, Injection, Incubation, Staining

    Binding of HA with GP2 is mediated by the carbohydrate-binding activities of HA. ( a , b ) Alexa Fluor 568-labelled HA subcomponents or reconstituted HA (Alexa Fluor 568-labelled HA2/HA3 or HA1/Alexa Fluor 568-labelled HA2/HA3; red) were injected into ligated mouse intestinal loops and incubated for 2 h; FAE regions were stained with FITC-labelled UEA-1 (green). ( c ) Recombinant Fc proteins (mGP2 and hGP2) were incubated with Strep-Tactin Superflow agarose pre-bound to HA1, HA3, HA2/HA3 core complex or HA (HA1/HA2/HA3). HA-bound proteins were analyzed by immunoblotting using HRP-labelled anti-human IgG antibody. ( d ) Reconstituted WT HA (HA1/Alexa Fluor 568-labelled HA2/HA3), mutant HA complex harbouring mutant HA1 (N285A, HA1N285A/Alexa Fluor 568-labelled HA2/HA3), mutant HA complex harbouring mutant HA3 (R528A, HA1/Alexa Fluor 568-labelled HA2/HA3R528A) and mutant HA complex harbouring mutant HA1 and HA3 (N285A/R528A, HA1N285A/Alexa Fluor 568-labelled HA2/HA3R528A; red) were injected into ligated mouse intestinal loops and incubated for 2 h; FAE regions were stained with FITC-labelled UEA-1 (green). ( e ) Recombinant mGP2-Fc protein was incubated with Strep-Tactin Superflow agarose pre-bound to HA (WT, N285A, R528A or N285A/R528A). HA-bound proteins were analyzed by immunoblotting using HRP-labelled anti-human IgG antibody. ( f ) GST–mGP2 and GST were incubated with Strep-Tactin Superflow agarose pre-bound to HA (WT). HA-bound proteins were analyzed by immunoblotting using anti-GST antibody and HRP-labelled anti-rabbit IgG antibody. Scale bar, 10 μm ( a , b , d ). Data are representative of two ( a , d ) or three ( b , c , e , f ) independent experiments.
    Figure Legend Snippet: Binding of HA with GP2 is mediated by the carbohydrate-binding activities of HA. ( a , b ) Alexa Fluor 568-labelled HA subcomponents or reconstituted HA (Alexa Fluor 568-labelled HA2/HA3 or HA1/Alexa Fluor 568-labelled HA2/HA3; red) were injected into ligated mouse intestinal loops and incubated for 2 h; FAE regions were stained with FITC-labelled UEA-1 (green). ( c ) Recombinant Fc proteins (mGP2 and hGP2) were incubated with Strep-Tactin Superflow agarose pre-bound to HA1, HA3, HA2/HA3 core complex or HA (HA1/HA2/HA3). HA-bound proteins were analyzed by immunoblotting using HRP-labelled anti-human IgG antibody. ( d ) Reconstituted WT HA (HA1/Alexa Fluor 568-labelled HA2/HA3), mutant HA complex harbouring mutant HA1 (N285A, HA1N285A/Alexa Fluor 568-labelled HA2/HA3), mutant HA complex harbouring mutant HA3 (R528A, HA1/Alexa Fluor 568-labelled HA2/HA3R528A) and mutant HA complex harbouring mutant HA1 and HA3 (N285A/R528A, HA1N285A/Alexa Fluor 568-labelled HA2/HA3R528A; red) were injected into ligated mouse intestinal loops and incubated for 2 h; FAE regions were stained with FITC-labelled UEA-1 (green). ( e ) Recombinant mGP2-Fc protein was incubated with Strep-Tactin Superflow agarose pre-bound to HA (WT, N285A, R528A or N285A/R528A). HA-bound proteins were analyzed by immunoblotting using HRP-labelled anti-human IgG antibody. ( f ) GST–mGP2 and GST were incubated with Strep-Tactin Superflow agarose pre-bound to HA (WT). HA-bound proteins were analyzed by immunoblotting using anti-GST antibody and HRP-labelled anti-rabbit IgG antibody. Scale bar, 10 μm ( a , b , d ). Data are representative of two ( a , d ) or three ( b , c , e , f ) independent experiments.

    Techniques Used: Binding Assay, Injection, Incubation, Staining, Recombinant, Mutagenesis

    L-PTC is taken up by M cells via the GP2–HA interaction. ( a ) Gp2 +/+ or Gp2 −/− mice were inoculated intragastrically (1.33 pmol: 1.0 μg) or intraperitoneally (0.067 fmol: 50 pg) with L-PTC (intragastrically , n =15 per group; intraperitoneally, n =5 per group). Statistical analyses were performed with the log-rank test. ( b ) Alexa Fluor 568-labelled L-PTC (red) were injected into ligated mouse intestinal loops and incubated for 1 h. PPs removed from Gp2 +/+ or Gp2 −/− mice were incubated with FITC-labelled UEA-1 (green). Images from FAE regions were analyzed quantitatively using the MetaMorph software. The data are expressed as percentages of the level in Gp2 +/+ mice. Error bars indicate s.d. For each group, six different PPs obtained from three different mice were examined ( Gp2 +/+ , 127 cells; Gp2 −/− , 161 cells). Statistical analyses were performed with Mann–Whitney U -test. Scale bar, 10 μm. NS, not significant.
    Figure Legend Snippet: L-PTC is taken up by M cells via the GP2–HA interaction. ( a ) Gp2 +/+ or Gp2 −/− mice were inoculated intragastrically (1.33 pmol: 1.0 μg) or intraperitoneally (0.067 fmol: 50 pg) with L-PTC (intragastrically , n =15 per group; intraperitoneally, n =5 per group). Statistical analyses were performed with the log-rank test. ( b ) Alexa Fluor 568-labelled L-PTC (red) were injected into ligated mouse intestinal loops and incubated for 1 h. PPs removed from Gp2 +/+ or Gp2 −/− mice were incubated with FITC-labelled UEA-1 (green). Images from FAE regions were analyzed quantitatively using the MetaMorph software. The data are expressed as percentages of the level in Gp2 +/+ mice. Error bars indicate s.d. For each group, six different PPs obtained from three different mice were examined ( Gp2 +/+ , 127 cells; Gp2 −/− , 161 cells). Statistical analyses were performed with Mann–Whitney U -test. Scale bar, 10 μm. NS, not significant.

    Techniques Used: Mouse Assay, Injection, Incubation, Software, MANN-WHITNEY

    2) Product Images from "The eukaryotic translation initiation factor eIF4E harnesses hyaluronan production to drive its malignant activity"

    Article Title: The eukaryotic translation initiation factor eIF4E harnesses hyaluronan production to drive its malignant activity

    Journal: eLife

    doi: 10.7554/eLife.29830

    eIF4E overexpression correlates with increased HA synthesis. ( A ) Fluorescence staining of HA (in green) using biotinylated HA-binding protein with streptavidin-FITC in U2Os cells overexpressing eIF4E, S53A mutant or vector control in the presence or absence of Streptomyces Hyaluronidase treatment. DAPI is in blue. Note cell surface expression of HA in response to eIF4E overexpression. All confocal settings are identical between specimens and thus lower signal is indicative of less HA. A × 40 objective with no digital zoom was used. ( B ) 2x digital zoom in confocal images of HA from part ( A ). ( C ) Quantification of fluorophore-assisted carbohydrate electrophoresis (FACE) gels (Sup Figure 1e f ) for HA levels in U2Os cells expressing eIF4E, S53A mutant or vector control, and U2Os cells overexpressing eIF4E following HAS3/eIF4E knockdown or pharmacological inhibition with ribavirin (Rib). ( D ) Fluorescence staining of HA (in green) following siRNA to eIF4E or ribavirin treatment in U2Os cells overexpressing eIF4E. DAPI is in blue. A × 63 objective with no digital zoom used. For bar graphs, the mean ± SD are shown. Experiments were carried out in triplicate, at least three independent times. **p
    Figure Legend Snippet: eIF4E overexpression correlates with increased HA synthesis. ( A ) Fluorescence staining of HA (in green) using biotinylated HA-binding protein with streptavidin-FITC in U2Os cells overexpressing eIF4E, S53A mutant or vector control in the presence or absence of Streptomyces Hyaluronidase treatment. DAPI is in blue. Note cell surface expression of HA in response to eIF4E overexpression. All confocal settings are identical between specimens and thus lower signal is indicative of less HA. A × 40 objective with no digital zoom was used. ( B ) 2x digital zoom in confocal images of HA from part ( A ). ( C ) Quantification of fluorophore-assisted carbohydrate electrophoresis (FACE) gels (Sup Figure 1e f ) for HA levels in U2Os cells expressing eIF4E, S53A mutant or vector control, and U2Os cells overexpressing eIF4E following HAS3/eIF4E knockdown or pharmacological inhibition with ribavirin (Rib). ( D ) Fluorescence staining of HA (in green) following siRNA to eIF4E or ribavirin treatment in U2Os cells overexpressing eIF4E. DAPI is in blue. A × 63 objective with no digital zoom used. For bar graphs, the mean ± SD are shown. Experiments were carried out in triplicate, at least three independent times. **p

    Techniques Used: Over Expression, Fluorescence, Staining, Binding Assay, Mutagenesis, Plasmid Preparation, Expressing, Electrophoresis, Inhibition

    3) Product Images from "Herpes Simplex Virus Type 1 Infection Induces Activation and Recruitment of Protein Kinase C to the Nuclear Membrane and Increased Phosphorylation of Lamin B"

    Article Title: Herpes Simplex Virus Type 1 Infection Induces Activation and Recruitment of Protein Kinase C to the Nuclear Membrane and Increased Phosphorylation of Lamin B

    Journal: Journal of Virology

    doi: 10.1128/JVI.80.1.494-504.2006

    Digital images of confocal fluorescence micrographs of mock-infected HEp-2 cells (A to C) or HEp-2 cells infected with HSV-1 (F) (D to F), R7037 U S 3-null HSV-1 mutant (G to I), or vRR1204 U S 3-kinase-defective HSV-1 mutant (J to L). At 16 h postinfection, cells were fixed and immunostained for PKC (FITC; green channel), lamin B (Texas Red; red channel), and ICP4 (Cy5; blue channel).
    Figure Legend Snippet: Digital images of confocal fluorescence micrographs of mock-infected HEp-2 cells (A to C) or HEp-2 cells infected with HSV-1 (F) (D to F), R7037 U S 3-null HSV-1 mutant (G to I), or vRR1204 U S 3-kinase-defective HSV-1 mutant (J to L). At 16 h postinfection, cells were fixed and immunostained for PKC (FITC; green channel), lamin B (Texas Red; red channel), and ICP4 (Cy5; blue channel).

    Techniques Used: Fluorescence, Infection, Mutagenesis

    Time course of PKC upregulation and recruitment to the nuclear rim. Digital images of confocal fluorescence micrographs were taken of HEp-2 cells infected with HSV-1 (F). At 0 h (A to C), 4 h (D to F), 8 h (G to I), 12 h (J to L), and 16 h (M to O) postinfection, cells were fixed and stained for PKC (FITC; green channel), lamin B (Texas Red; red channel), and ICP4 (Cy5; blue channel).
    Figure Legend Snippet: Time course of PKC upregulation and recruitment to the nuclear rim. Digital images of confocal fluorescence micrographs were taken of HEp-2 cells infected with HSV-1 (F). At 0 h (A to C), 4 h (D to F), 8 h (G to I), 12 h (J to L), and 16 h (M to O) postinfection, cells were fixed and stained for PKC (FITC; green channel), lamin B (Texas Red; red channel), and ICP4 (Cy5; blue channel).

    Techniques Used: Fluorescence, Infection, Staining

    Digital images of confocal fluorescence micrographs of mock-infected (A to D and I to L) and HSV-1 (F)-infected (E to H and M to P) HEp-2 cells (A to H) and HeLa cells (I to P). Cells were fixed in methanol at 16 h postinfection and incubated with anti-PKC, anti-lamin B, and anti-ICP4 antibody. Anti-PKC antibody was stained with FITC-conjugated secondary antibody (green channel), anti-lamin B antibody was stained with Texas Red-conjugated secondary antibody (red channel), and ICP4 was stained with Cy5-conjugated secondary antibody (blue channel).
    Figure Legend Snippet: Digital images of confocal fluorescence micrographs of mock-infected (A to D and I to L) and HSV-1 (F)-infected (E to H and M to P) HEp-2 cells (A to H) and HeLa cells (I to P). Cells were fixed in methanol at 16 h postinfection and incubated with anti-PKC, anti-lamin B, and anti-ICP4 antibody. Anti-PKC antibody was stained with FITC-conjugated secondary antibody (green channel), anti-lamin B antibody was stained with Texas Red-conjugated secondary antibody (red channel), and ICP4 was stained with Cy5-conjugated secondary antibody (blue channel).

    Techniques Used: Fluorescence, Infection, Incubation, Staining

    Digital confocal images of mock-infected or HSV-1 (F)-infected HEp-2 cells immunostained with PKC-specific antibodies. At 16 h postinfection, cells were fixed with methanol and incubated with anti-lamin B antibody, anti-ICP4 antibody, and one of the following PKC-specific antibodies: anti-PKCα (A to D), anti-PKCδ (E to H), anti-PKCζ (I to L). Anti-PKC antibodies were stained with FITC-conjugated secondary antibody (green channel), anti-lamin B antibody was stained with Texas Red-conjugated secondary antibody (red channel), and ICP4 was stained with Cy5-conjugated secondary antibody (blue channel).
    Figure Legend Snippet: Digital confocal images of mock-infected or HSV-1 (F)-infected HEp-2 cells immunostained with PKC-specific antibodies. At 16 h postinfection, cells were fixed with methanol and incubated with anti-lamin B antibody, anti-ICP4 antibody, and one of the following PKC-specific antibodies: anti-PKCα (A to D), anti-PKCδ (E to H), anti-PKCζ (I to L). Anti-PKC antibodies were stained with FITC-conjugated secondary antibody (green channel), anti-lamin B antibody was stained with Texas Red-conjugated secondary antibody (red channel), and ICP4 was stained with Cy5-conjugated secondary antibody (blue channel).

    Techniques Used: Infection, Incubation, Staining

    Effects of U L 31 and U L 34 and their association on HSV-1-induced PKC recruitment. Digital confocal images show mock-infected HEp-2 cells (A to C) or HEp-2 cells infected with HSV-1 (F) (D to F), v3161 U L 31-null HSV-1 mutant (G to I), vRR1072 U L 34-null HSV-1 mutant (J to L), or v3480 U L 34-U L 31 association-defective mutant (M to O). At 16 h postinfection, cells were fixed and stained for PKC (FITC; green channel), lamin B (Texas Red; red channel), and ICP4 (Cy5; blue channel).
    Figure Legend Snippet: Effects of U L 31 and U L 34 and their association on HSV-1-induced PKC recruitment. Digital confocal images show mock-infected HEp-2 cells (A to C) or HEp-2 cells infected with HSV-1 (F) (D to F), v3161 U L 31-null HSV-1 mutant (G to I), vRR1072 U L 34-null HSV-1 mutant (J to L), or v3480 U L 34-U L 31 association-defective mutant (M to O). At 16 h postinfection, cells were fixed and stained for PKC (FITC; green channel), lamin B (Texas Red; red channel), and ICP4 (Cy5; blue channel).

    Techniques Used: Infection, Mutagenesis, Staining

    Digital images of confocal fluorescence micrographs of HSV-1 (F)-infected HeLa cells. At 16 h postinfection, cells were fixed and immunostained for lamin B (Texas Red; red channel), lamin A (FITC; green channel), and ICP4 (Cy5; blue channel).
    Figure Legend Snippet: Digital images of confocal fluorescence micrographs of HSV-1 (F)-infected HeLa cells. At 16 h postinfection, cells were fixed and immunostained for lamin B (Texas Red; red channel), lamin A (FITC; green channel), and ICP4 (Cy5; blue channel).

    Techniques Used: Fluorescence, Infection

    4) Product Images from "The Role of cis Dimerization of Signal Regulatory Protein ? (SIRP?) in Binding to CD47 *"

    Article Title: The Role of cis Dimerization of Signal Regulatory Protein ? (SIRP?) in Binding to CD47 *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.180018

    SIRPα dimerization is disrupted after treatment with tunicamycin. A , flow cytometric analysis of tunicamycin-treated ( solid line ) and untreated ( dashed line ) CHO-SIRPα transfectants labeled with PHA-L-FITC ( top left panel ) to measure the
    Figure Legend Snippet: SIRPα dimerization is disrupted after treatment with tunicamycin. A , flow cytometric analysis of tunicamycin-treated ( solid line ) and untreated ( dashed line ) CHO-SIRPα transfectants labeled with PHA-L-FITC ( top left panel ) to measure the

    Techniques Used: Flow Cytometry, Labeling

    5) Product Images from "Mechanical force-driven TNFα endocytosis governs stem cell homeostasis"

    Article Title: Mechanical force-driven TNFα endocytosis governs stem cell homeostasis

    Journal: Bone Research

    doi: 10.1038/s41413-020-00117-x

    Mechanical force drives TNFα endocytosis and promotes MSC function. a , b Examination of secreted concentrations and protein expression levels of TNFα. MSCs underwent cyclic loading of stretch force at 15% elongation, 0.5 Hz. N = 3. c Western blot analyses of mTOR signaling ( N = 3). d – f Functional analyses of MSCs according to BrdU labeling and osteogenic and adipogenic differentiation. N = 3. Scale bars = 100 μm. Stretch force was applied twice per week for 2 h per session. g qRT-PCR analysis of the mRNA expression levels of TNFα ( N = 3). h , i Endocytosis analysis of FITC-labeled TNFα uptake by MSCs for 2 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . Stretch force was applied for a 2 h period. N = 3. Scale bars = 20 μm. j , k Endocytosis analysis of Alexa Fluor TM 488-conjugated bovine serum albumin (488-BSA) taken up by MSCs for 2 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . Stretch force was applied for a 2 h period. N = 3. Scale bars = 20 μm. l Diagram showing mechanical force-driven TNFα endocytosis. For quantification of Western blotting, a two-tailed Student’s t test was used for the comparison between the treatment and stretch 0 h/control groups. * P
    Figure Legend Snippet: Mechanical force drives TNFα endocytosis and promotes MSC function. a , b Examination of secreted concentrations and protein expression levels of TNFα. MSCs underwent cyclic loading of stretch force at 15% elongation, 0.5 Hz. N = 3. c Western blot analyses of mTOR signaling ( N = 3). d – f Functional analyses of MSCs according to BrdU labeling and osteogenic and adipogenic differentiation. N = 3. Scale bars = 100 μm. Stretch force was applied twice per week for 2 h per session. g qRT-PCR analysis of the mRNA expression levels of TNFα ( N = 3). h , i Endocytosis analysis of FITC-labeled TNFα uptake by MSCs for 2 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . Stretch force was applied for a 2 h period. N = 3. Scale bars = 20 μm. j , k Endocytosis analysis of Alexa Fluor TM 488-conjugated bovine serum albumin (488-BSA) taken up by MSCs for 2 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . Stretch force was applied for a 2 h period. N = 3. Scale bars = 20 μm. l Diagram showing mechanical force-driven TNFα endocytosis. For quantification of Western blotting, a two-tailed Student’s t test was used for the comparison between the treatment and stretch 0 h/control groups. * P

    Techniques Used: Expressing, Western Blot, Functional Assay, Labeling, Quantitative RT-PCR, In Vitro, Two Tailed Test

    TNFα safeguards MSC homeostasis in a receptor-independent manner through endocytosis. a – d Functional analyses of MSCs according to CFU, BrdU labeling, and osteogenic and adipogenic differentiation. MSCs were derived from WT, TNFα −/− or TNFR −/− mice ( N = 5). Scale bars = 100 μm. e Calcein labeling for bone formation analysis in WT and TNFR −/− mice ( N = 5). Scale bar = 50 μm. f Oli red O staining for bone marrow adiposity in WT and TNFR −/− mice ( N = 5). Scale bar = 150 μm. g Endocytosis analysis of FITC-labeled TNFα uptake by MSCs for 24 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . N = 3. Scale bars = 20 μm (top) and 7 μm (bottom). h Western blot analysis of protein expression levels ( N = 3). i – k Functional analyses of MSCs according to BrdU labeling and osteogenic and adipogenic differentiation. N = 3. Scale bars = 100 μm. l Diagram showing that TNFα regulates MSC homeostasis in a receptor-independent manner through endocytosis. For quantification of Western blotting, a two-tailed Student’s t test was used for the comparison between the treatment and WT groups. * P
    Figure Legend Snippet: TNFα safeguards MSC homeostasis in a receptor-independent manner through endocytosis. a – d Functional analyses of MSCs according to CFU, BrdU labeling, and osteogenic and adipogenic differentiation. MSCs were derived from WT, TNFα −/− or TNFR −/− mice ( N = 5). Scale bars = 100 μm. e Calcein labeling for bone formation analysis in WT and TNFR −/− mice ( N = 5). Scale bar = 50 μm. f Oli red O staining for bone marrow adiposity in WT and TNFR −/− mice ( N = 5). Scale bar = 150 μm. g Endocytosis analysis of FITC-labeled TNFα uptake by MSCs for 24 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . N = 3. Scale bars = 20 μm (top) and 7 μm (bottom). h Western blot analysis of protein expression levels ( N = 3). i – k Functional analyses of MSCs according to BrdU labeling and osteogenic and adipogenic differentiation. N = 3. Scale bars = 100 μm. l Diagram showing that TNFα regulates MSC homeostasis in a receptor-independent manner through endocytosis. For quantification of Western blotting, a two-tailed Student’s t test was used for the comparison between the treatment and WT groups. * P

    Techniques Used: Functional Assay, Labeling, Derivative Assay, Mouse Assay, Staining, In Vitro, Western Blot, Expressing, Two Tailed Test

    6) Product Images from "Decoding Single Cell Morphology in Osteotropic Breast Cancer Cells for Dissecting Their Migratory, Molecular and Biophysical Heterogeneity"

    Article Title: Decoding Single Cell Morphology in Osteotropic Breast Cancer Cells for Dissecting Their Migratory, Molecular and Biophysical Heterogeneity

    Journal: Cancers

    doi: 10.3390/cancers14030603

    Differential binding of lectins to breast cancer cell lines. ( A , B ) The parental cells MB-231 and its bone-seeking derivatives, MET and BONE, were cell-surface labeled with one of a panel of distinct FITC-conjugated lectins prior to cytochemistry (( A ), left panels) and flow cytometry (( A ), right panels, ( B )) analyses. See also Figure S4 . Percentages of positive cells are indicated in the histograms ( A ) and the median fluorescence intensity is presented for each lectin ( B ). Data from representative experiments acquired under uniform instruments setting for all cell lines are displayed. The lectins used were Concanavalin A (ConA), Datura Stramonium Lectin (DSL), Dolichos Biflorus Agglutinin (DBA), Erythrina Cristagalli Lectin (ECL), Griffonia Simplicifolia Lectin I (GSL-I), Griffonia Simplicifolia Lectin II (GSL-II), Lens Culinaris Agglutinin (LCA), Lycopersicon Esculentum Lectin (LEL), Peanut Agglutinin (PNA), Phaseolus Vulgaris Lectin E (PHA-E), Phaseolus Vulgaris Lectin L (PHA-L), Pisum Sativum Agglutinin (PSA), Ricinus Communis Agglutinin I (RCA), Sambucus Nigra Lectin (SNA), Solanum Tuberosum Lectin (STL), Soybean Agglutinin (SBA), Ulex Europaeus Agglutinin I (UEA-I), Vicia Villosa Lectin (VVL) and Wheat Germ Agglutinin (WGA, succinylated (succ)-WGA). Scale bar, 25 μm.
    Figure Legend Snippet: Differential binding of lectins to breast cancer cell lines. ( A , B ) The parental cells MB-231 and its bone-seeking derivatives, MET and BONE, were cell-surface labeled with one of a panel of distinct FITC-conjugated lectins prior to cytochemistry (( A ), left panels) and flow cytometry (( A ), right panels, ( B )) analyses. See also Figure S4 . Percentages of positive cells are indicated in the histograms ( A ) and the median fluorescence intensity is presented for each lectin ( B ). Data from representative experiments acquired under uniform instruments setting for all cell lines are displayed. The lectins used were Concanavalin A (ConA), Datura Stramonium Lectin (DSL), Dolichos Biflorus Agglutinin (DBA), Erythrina Cristagalli Lectin (ECL), Griffonia Simplicifolia Lectin I (GSL-I), Griffonia Simplicifolia Lectin II (GSL-II), Lens Culinaris Agglutinin (LCA), Lycopersicon Esculentum Lectin (LEL), Peanut Agglutinin (PNA), Phaseolus Vulgaris Lectin E (PHA-E), Phaseolus Vulgaris Lectin L (PHA-L), Pisum Sativum Agglutinin (PSA), Ricinus Communis Agglutinin I (RCA), Sambucus Nigra Lectin (SNA), Solanum Tuberosum Lectin (STL), Soybean Agglutinin (SBA), Ulex Europaeus Agglutinin I (UEA-I), Vicia Villosa Lectin (VVL) and Wheat Germ Agglutinin (WGA, succinylated (succ)-WGA). Scale bar, 25 μm.

    Techniques Used: Binding Assay, Labeling, Flow Cytometry, Fluorescence, Whole Genome Amplification

    7) Product Images from "Passive Immunization Reduces Behavioral and Neuropathological Deficits in an Alpha-Synuclein Transgenic Model of Lewy Body Disease"

    Article Title: Passive Immunization Reduces Behavioral and Neuropathological Deficits in an Alpha-Synuclein Transgenic Model of Lewy Body Disease

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0019338

    Co-localization of the FITC-tagged α-syn 9E4 antibody with lysosomal and autophagosomal markers. To analyze the sub-cellular distribution of the 9E4 antibody immunohistochemical and ultrastructural analysis was conducted in 9E4-FITC immunized α-syn tg mice. (A–C) Representative confocal image of a brain section from an α-syn tg mouse immunized 9E4-FITC and co-labeled with an antibody against α-syn. Arrows indicate co-localization of the 9E4-FITC signal with α-syn in neuronal granular-like structures. (D–F) Confocal image from an α-syn tg mouse immunized 9E4-FITC and co-labeled with an antibody against LC3. Arrows indicate co-localization of the 9E4-FITC with LC3 in neuronal autophagosome-like structures. (G–I) Confocal image from an α-syn tg mouse immunized 9E4-FITC and co-labeled with an antibody against cathepsin D. Arrows indicate co-localization of the 9E4-FITC with cathepsin-D in neuronal lysosomal-like structuresLC3. (J, K) Representative electron micrographs of sections from an α-syn tg mouse immunized with the 9E4 antibody and immunolabeld with gold-tagged anti-mouse antibody. (L, M) Electron micrographs of sections from an α-syn tg mouse immunized with the control IgG1 antibody and immunolabeld with gold-tagged anti-mouse antibody. (N, O) Representative electron micrographs of sections from a non-tg mouse immunized with the 9E4 antibody and immunolabeld with gold-tagged anti-mouse antibody. No reactivity is observed in lysosomes or autophagosomes. Scale bar (A–I) = 10 µM; (J–O) magnification 25,000x.
    Figure Legend Snippet: Co-localization of the FITC-tagged α-syn 9E4 antibody with lysosomal and autophagosomal markers. To analyze the sub-cellular distribution of the 9E4 antibody immunohistochemical and ultrastructural analysis was conducted in 9E4-FITC immunized α-syn tg mice. (A–C) Representative confocal image of a brain section from an α-syn tg mouse immunized 9E4-FITC and co-labeled with an antibody against α-syn. Arrows indicate co-localization of the 9E4-FITC signal with α-syn in neuronal granular-like structures. (D–F) Confocal image from an α-syn tg mouse immunized 9E4-FITC and co-labeled with an antibody against LC3. Arrows indicate co-localization of the 9E4-FITC with LC3 in neuronal autophagosome-like structures. (G–I) Confocal image from an α-syn tg mouse immunized 9E4-FITC and co-labeled with an antibody against cathepsin D. Arrows indicate co-localization of the 9E4-FITC with cathepsin-D in neuronal lysosomal-like structuresLC3. (J, K) Representative electron micrographs of sections from an α-syn tg mouse immunized with the 9E4 antibody and immunolabeld with gold-tagged anti-mouse antibody. (L, M) Electron micrographs of sections from an α-syn tg mouse immunized with the control IgG1 antibody and immunolabeld with gold-tagged anti-mouse antibody. (N, O) Representative electron micrographs of sections from a non-tg mouse immunized with the 9E4 antibody and immunolabeld with gold-tagged anti-mouse antibody. No reactivity is observed in lysosomes or autophagosomes. Scale bar (A–I) = 10 µM; (J–O) magnification 25,000x.

    Techniques Used: Immunohistochemistry, Mouse Assay, Labeling

    Trafficking of the FITC-tagged α-syn 9E4 antibody in tg mice. To investigate the distribution of the 9E4 antibody after passive immunization, the FITC tagged antibody was injected intravenously (IV) and analyzed by ELISA and confocal microscopy. (A) Antibody titers in the plasma and brain at 3, 14 and 30 days post-injection in mice immunized with the 9E4 antibody, determined by ELISA. (B) Image analysis of 9E4-FITC positive neurons in the α-syn tg mice at 3, 14 and 30 days post-injection. (C, D) Representative laser scanning confocal images of the signal in the FITC channel in the temporal cortex of α-syn tg mouse 30 days following intravenous IV injection with the FITC-tagged 9E4 antibody. Arrows highlight labeled intra-neuronal granular-like structures. (E) No signal is detected in the FITC channel in α-syn tg mouse 30 days following IV injection with the FITC-tagged IgG1 control antibody. (F) No signal in the FITC channel in non-tg mouse 30 days following IV injection with the FITC-tagged 9E4 antibody. (G) Confocal image of a section from an antibody-naive α-syn tg mouse immunolabeled with cerebrospinal fluid (CSF) from a mouse immunized with 9E4-FITC. (H) Confocal image of a section from non-tg mouse immunolabeled 9E4-FITC antibody. Scale bar (C, E–H) = 50 µM; (D) = 10 µM. N = 20 per group, 12 months of age. Error bars represent mean ± SEM.
    Figure Legend Snippet: Trafficking of the FITC-tagged α-syn 9E4 antibody in tg mice. To investigate the distribution of the 9E4 antibody after passive immunization, the FITC tagged antibody was injected intravenously (IV) and analyzed by ELISA and confocal microscopy. (A) Antibody titers in the plasma and brain at 3, 14 and 30 days post-injection in mice immunized with the 9E4 antibody, determined by ELISA. (B) Image analysis of 9E4-FITC positive neurons in the α-syn tg mice at 3, 14 and 30 days post-injection. (C, D) Representative laser scanning confocal images of the signal in the FITC channel in the temporal cortex of α-syn tg mouse 30 days following intravenous IV injection with the FITC-tagged 9E4 antibody. Arrows highlight labeled intra-neuronal granular-like structures. (E) No signal is detected in the FITC channel in α-syn tg mouse 30 days following IV injection with the FITC-tagged IgG1 control antibody. (F) No signal in the FITC channel in non-tg mouse 30 days following IV injection with the FITC-tagged 9E4 antibody. (G) Confocal image of a section from an antibody-naive α-syn tg mouse immunolabeled with cerebrospinal fluid (CSF) from a mouse immunized with 9E4-FITC. (H) Confocal image of a section from non-tg mouse immunolabeled 9E4-FITC antibody. Scale bar (C, E–H) = 50 µM; (D) = 10 µM. N = 20 per group, 12 months of age. Error bars represent mean ± SEM.

    Techniques Used: Mouse Assay, Injection, Enzyme-linked Immunosorbent Assay, Confocal Microscopy, IV Injection, Labeling, Immunolabeling

    8) Product Images from "Vibropolyfection: coupling polymer-mediated gene delivery to mechanical stimulation to enhance transfection of adherent cells"

    Article Title: Vibropolyfection: coupling polymer-mediated gene delivery to mechanical stimulation to enhance transfection of adherent cells

    Journal: Journal of Nanobiotechnology

    doi: 10.1186/s12951-022-01571-x

    Effect of vibrational loading on polyplex uptake and transgene expression. a Fold-increase in pDNA uptake by polyfected vs. vibropolyfected L929 cells. Experiments were carried out with l PEI/pGLuc following the post-delivery set-up. Results express the fold-increase in the amount of intracellular pDNA (t = 24 hrs) in vibropolyfected cells with respect to polyfected counterparts. Results are expressed as mean ± SD (n = 3). b Expression levels of Gaussia luciferase transcripts in L929 cells polyfected and vibropolyfected with l PEI/pGLuc. Target luciferase transcript levels were normalized to GAPDH levels. Results are expressed as mean ± SD (n = 3). c Uptake kinetics of l PEI/pDNA-FITC polyplexes. CLSM images of L929 cells transfected according to standard transfection vs. vibropolyfection (post-delivery set-up, 1,000 Hz-vibrational loading for 5 min). Cells were next fixed and imaged at 30 min, 1 hr, and 4 hrs post-polyplex addition to the cells. High-magnification (insets of c , right panels) digital images of single cells were taken 4 hrs after polyfection or vibropolyfection (post-delivery set-up, 1,000 Hz-vibrational loading for 5 min). Polyplexes are in green, while F-actin and nuclei are in red and blue, respectively. Scale bars = 50 µm and 12.5 µm (right panels)
    Figure Legend Snippet: Effect of vibrational loading on polyplex uptake and transgene expression. a Fold-increase in pDNA uptake by polyfected vs. vibropolyfected L929 cells. Experiments were carried out with l PEI/pGLuc following the post-delivery set-up. Results express the fold-increase in the amount of intracellular pDNA (t = 24 hrs) in vibropolyfected cells with respect to polyfected counterparts. Results are expressed as mean ± SD (n = 3). b Expression levels of Gaussia luciferase transcripts in L929 cells polyfected and vibropolyfected with l PEI/pGLuc. Target luciferase transcript levels were normalized to GAPDH levels. Results are expressed as mean ± SD (n = 3). c Uptake kinetics of l PEI/pDNA-FITC polyplexes. CLSM images of L929 cells transfected according to standard transfection vs. vibropolyfection (post-delivery set-up, 1,000 Hz-vibrational loading for 5 min). Cells were next fixed and imaged at 30 min, 1 hr, and 4 hrs post-polyplex addition to the cells. High-magnification (insets of c , right panels) digital images of single cells were taken 4 hrs after polyfection or vibropolyfection (post-delivery set-up, 1,000 Hz-vibrational loading for 5 min). Polyplexes are in green, while F-actin and nuclei are in red and blue, respectively. Scale bars = 50 µm and 12.5 µm (right panels)

    Techniques Used: Expressing, Luciferase, Confocal Laser Scanning Microscopy, Transfection

    9) Product Images from "Subsets of Transgenic T Cells That Recognize CD1 Induce or Prevent Murine Lupus: Role of Cytokines "

    Article Title: Subsets of Transgenic T Cells That Recognize CD1 Induce or Prevent Murine Lupus: Role of Cytokines

    Journal: The Journal of Experimental Medicine

    doi:

    Immunohistopathology of kidneys from BALB/c nu/nu recipients given transgenic BM cells. ( A and B) Two-color flow cytometric analysis of enriched T cells from the BM of SP or DN transgenic mice, respectively, stained for CD4 and CD8 receptors versus V β 9 receptors. Upper boxes enclose CD4 + and CD8 + T cells and lower box encloses CD4 − CD8 − T cells. T cells were enriched by depleting B220 + , Gr-1 + , and Mac-1 + cells ( 16 ). ( C and D ) The immunofluorescent staining of histological sections of kidneys from recipients given BM cells from SP or DN transgenic mice, respectively, with FITC-conjugated anti–mouse IgG antibodies. Arrows show staining of capillary loops. E and F show the appearance of recipients given BM cells from SP or DN transgenic mice, respectively. The mouse in E has ascites and generalized edema.
    Figure Legend Snippet: Immunohistopathology of kidneys from BALB/c nu/nu recipients given transgenic BM cells. ( A and B) Two-color flow cytometric analysis of enriched T cells from the BM of SP or DN transgenic mice, respectively, stained for CD4 and CD8 receptors versus V β 9 receptors. Upper boxes enclose CD4 + and CD8 + T cells and lower box encloses CD4 − CD8 − T cells. T cells were enriched by depleting B220 + , Gr-1 + , and Mac-1 + cells ( 16 ). ( C and D ) The immunofluorescent staining of histological sections of kidneys from recipients given BM cells from SP or DN transgenic mice, respectively, with FITC-conjugated anti–mouse IgG antibodies. Arrows show staining of capillary loops. E and F show the appearance of recipients given BM cells from SP or DN transgenic mice, respectively. The mouse in E has ascites and generalized edema.

    Techniques Used: Transgenic Assay, Flow Cytometry, Mouse Assay, Staining

    10) Product Images from "Toxoplasma gondii AP2IX-4 regulates gene expression during bradyzoite development"

    Article Title: Toxoplasma gondii AP2IX-4 regulates gene expression during bradyzoite development

    Journal: bioRxiv

    doi: 10.1101/104208

    Expression of AP2IX-4 in developing bradyzoites in vitro . A. IFAs were performed on Δap2IX-4 ::AP2IX-4 HA parasites during tachyzoite and developing bradyzoite stages up to 8 days in alkaline pH. FITC-conjugated Dolichos lectin was used to visualize bradyzoite cyst walls (the LDH2-GFP reporter also appears in bradyzoites in this channel). AP2IX-4 HA protein was visualized using anti-HA (red) and DAPI (blue) was used to stain DNA. The percent of parasites expressing AP2IX-4 HA within 50 random vacuoles was recorded (%HA+), noting that it was not possible to accurately discern the total number of parasites in tissue cysts beyond 4 days post-induction (ND = not determined). Values represent the average and standard deviation for 3 independent experiments. B. IFAs performed with anti-IMC3 antibody (yellow) on Δap2IX-4 ::AP2IX-4 HA parasites following 4 days in alkaline pH. The LDH2-GFP reporter (green) identifies bradyzoites. AP2IX-4 HA protein was visualized using anti-HA (red). Arrowheads point to budding daughter cells in dividing parasites expressing AP2IX-4 HA . Scale bar = 3 microns.
    Figure Legend Snippet: Expression of AP2IX-4 in developing bradyzoites in vitro . A. IFAs were performed on Δap2IX-4 ::AP2IX-4 HA parasites during tachyzoite and developing bradyzoite stages up to 8 days in alkaline pH. FITC-conjugated Dolichos lectin was used to visualize bradyzoite cyst walls (the LDH2-GFP reporter also appears in bradyzoites in this channel). AP2IX-4 HA protein was visualized using anti-HA (red) and DAPI (blue) was used to stain DNA. The percent of parasites expressing AP2IX-4 HA within 50 random vacuoles was recorded (%HA+), noting that it was not possible to accurately discern the total number of parasites in tissue cysts beyond 4 days post-induction (ND = not determined). Values represent the average and standard deviation for 3 independent experiments. B. IFAs performed with anti-IMC3 antibody (yellow) on Δap2IX-4 ::AP2IX-4 HA parasites following 4 days in alkaline pH. The LDH2-GFP reporter (green) identifies bradyzoites. AP2IX-4 HA protein was visualized using anti-HA (red). Arrowheads point to budding daughter cells in dividing parasites expressing AP2IX-4 HA . Scale bar = 3 microns.

    Techniques Used: Expressing, In Vitro, Staining, Standard Deviation

    11) Product Images from "Interaction between Bluetongue virus outer capsid protein VP2 and vimentin is necessary for virus egress"

    Article Title: Interaction between Bluetongue virus outer capsid protein VP2 and vimentin is necessary for virus egress

    Journal: Virology Journal

    doi: 10.1186/1743-422X-4-7

    Distribution of VP2 within infected and transfected Vero cells by fluorescence microscopy . A) Cells infected with BTV-10, B-E) transfected cells expressing B) VP2, C) VP2-GFP, D) GFP-VP2 and E) GFP only. VP2 in A and B were detected with anti VP2 monoclonal antibody (rabbit) and either FITC (A) or TRITC (B) conjugated secondary antibody. C-E were visualised based on GFP fluorescence. Expression of full-length, tagged VP2 variants was confirmed by western blot using an anti-GFP antibody (F).
    Figure Legend Snippet: Distribution of VP2 within infected and transfected Vero cells by fluorescence microscopy . A) Cells infected with BTV-10, B-E) transfected cells expressing B) VP2, C) VP2-GFP, D) GFP-VP2 and E) GFP only. VP2 in A and B were detected with anti VP2 monoclonal antibody (rabbit) and either FITC (A) or TRITC (B) conjugated secondary antibody. C-E were visualised based on GFP fluorescence. Expression of full-length, tagged VP2 variants was confirmed by western blot using an anti-GFP antibody (F).

    Techniques Used: Infection, Transfection, Fluorescence, Microscopy, Expressing, Western Blot

    12) Product Images from "Cerebral Neovascularization and Remodeling Patterns in Two Different Models of Type 2 Diabetes"

    Article Title: Cerebral Neovascularization and Remodeling Patterns in Two Different Models of Type 2 Diabetes

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0056264

    Glycemic control prevents neovascularization in the GK model of diabetes. (A) Vascular density is significantly increased in both cortex and striatum of GK rats and this was prevented by metformin treatment started at the onset of diabetes. (B) Vascular volume and (C) vascular surface area are also increased in diabetes. (D) Both micro and macrovascular volume as well as surface area (E) are increased in GK rats as compared to control. (F) Diabetic vascular correlations are extremely disproportionate and the slopes of the two lines are significantly different. (G, H) Immature cerebral microvessels are more abundant in diabetes. Representative images of cerebral striatal vasculature showing perfused (green-FITC dextran) and non-perfused vessels (Red-Isolectin). GK rats exhibit increased immature vasculature and glycemic control with metformin reduced the immature vasculature. F values are indicated under each ANOVA analysis group. *p
    Figure Legend Snippet: Glycemic control prevents neovascularization in the GK model of diabetes. (A) Vascular density is significantly increased in both cortex and striatum of GK rats and this was prevented by metformin treatment started at the onset of diabetes. (B) Vascular volume and (C) vascular surface area are also increased in diabetes. (D) Both micro and macrovascular volume as well as surface area (E) are increased in GK rats as compared to control. (F) Diabetic vascular correlations are extremely disproportionate and the slopes of the two lines are significantly different. (G, H) Immature cerebral microvessels are more abundant in diabetes. Representative images of cerebral striatal vasculature showing perfused (green-FITC dextran) and non-perfused vessels (Red-Isolectin). GK rats exhibit increased immature vasculature and glycemic control with metformin reduced the immature vasculature. F values are indicated under each ANOVA analysis group. *p

    Techniques Used:

    13) Product Images from "Flow Cytometry of Mouse and Human Adipocytes for the Analysis of Browning and Cellular Heterogeneity"

    Article Title: Flow Cytometry of Mouse and Human Adipocytes for the Analysis of Browning and Cellular Heterogeneity

    Journal: Cell Reports

    doi: 10.1016/j.celrep.2018.08.006

    Differential Sorting of Large and Small Adipocytes Using SSC as a Proxy for Adipocyte Size (A and B) FSC/SSC plots (A) showing the gates used to sort out SSC_high (small) and SSC_low (large) adipocytes from the same sample, as well as representative confocal images (B) of the sorted cells showing the cell outline stained with FITC-lectin (white). Scale bars, 20 μm. (C) Measured adipocyte diameters of cells sorted from the total scWAT sample (all cells) or sorted using the depicted SSC_high and SSC_low gates. Note that the SSC-based gates efficiently enrich for the occurrence of small or large cells, which are less common in the total sorted sample. Lines show means ± SDs. ∗∗∗∗ p
    Figure Legend Snippet: Differential Sorting of Large and Small Adipocytes Using SSC as a Proxy for Adipocyte Size (A and B) FSC/SSC plots (A) showing the gates used to sort out SSC_high (small) and SSC_low (large) adipocytes from the same sample, as well as representative confocal images (B) of the sorted cells showing the cell outline stained with FITC-lectin (white). Scale bars, 20 μm. (C) Measured adipocyte diameters of cells sorted from the total scWAT sample (all cells) or sorted using the depicted SSC_high and SSC_low gates. Note that the SSC-based gates efficiently enrich for the occurrence of small or large cells, which are less common in the total sorted sample. Lines show means ± SDs. ∗∗∗∗ p

    Techniques Used: Staining

    Developing the Adipocyte Flow Cytometry Protocol (A) Freshly isolated adipocytes and SVF cells from AdTomato and WT mice, stained with fluorescein isothiocyanate (FITC)-labeled lectin (green) and Hoechst (blue) and imaged for tdTomato expression (red). Scale bars, 100 μm; note the differences in length. (B) Flow cytometry using standard settings for scWAT adipocytes and samples containing only pure mouse lipid or sunflower seed oil. Adipocyte samples show ungated and tdTomato + (T + ) gated events. (C) Flow cytometry using optimized adipocyte flow cytometry settings for scWAT adipocytes, BAT adipocytes, or SVF cells, showing ungated and tdTomato + (T + ) or Hoechst + (H + ) gated events. .
    Figure Legend Snippet: Developing the Adipocyte Flow Cytometry Protocol (A) Freshly isolated adipocytes and SVF cells from AdTomato and WT mice, stained with fluorescein isothiocyanate (FITC)-labeled lectin (green) and Hoechst (blue) and imaged for tdTomato expression (red). Scale bars, 100 μm; note the differences in length. (B) Flow cytometry using standard settings for scWAT adipocytes and samples containing only pure mouse lipid or sunflower seed oil. Adipocyte samples show ungated and tdTomato + (T + ) gated events. (C) Flow cytometry using optimized adipocyte flow cytometry settings for scWAT adipocytes, BAT adipocytes, or SVF cells, showing ungated and tdTomato + (T + ) or Hoechst + (H + ) gated events. .

    Techniques Used: Flow Cytometry, Cytometry, Isolation, Mouse Assay, Staining, Labeling, Expressing

    14) Product Images from "Proliferation and Migration of Label-Retaining Cells of the Kidney Papilla"

    Article Title: Proliferation and Migration of Label-Retaining Cells of the Kidney Papilla

    Journal: Journal of the American Society of Nephrology : JASN

    doi: 10.1681/ASN.2008111203

    In vivo labeling and migration of renal papillary cells are shown. (A) Cells in the superficial papilla of rat were labeled. Rat kidney papilla were labeled with KPH26 and isolated 1 h later. Collecting ducts were labeled with Dolichos biflorus agglutinin-FITC
    Figure Legend Snippet: In vivo labeling and migration of renal papillary cells are shown. (A) Cells in the superficial papilla of rat were labeled. Rat kidney papilla were labeled with KPH26 and isolated 1 h later. Collecting ducts were labeled with Dolichos biflorus agglutinin-FITC

    Techniques Used: In Vivo, Labeling, Migration, Isolation

    Papillary cells labeled with dye after injury show upward migration. Labeled cells with dye were easily detected in the parenchyma of the papilla; collecting ducts were labeled with Dolichos biflorus agglutinin–FITC (top; bar = 100 μm).
    Figure Legend Snippet: Papillary cells labeled with dye after injury show upward migration. Labeled cells with dye were easily detected in the parenchyma of the papilla; collecting ducts were labeled with Dolichos biflorus agglutinin–FITC (top; bar = 100 μm).

    Techniques Used: Labeling, Migration

    15) Product Images from "Flow Cytometry of Mouse and Human Adipocytes for the Analysis of Browning and Cellular Heterogeneity"

    Article Title: Flow Cytometry of Mouse and Human Adipocytes for the Analysis of Browning and Cellular Heterogeneity

    Journal: Cell Reports

    doi: 10.1016/j.celrep.2018.08.006

    Differential Sorting of Large and Small Adipocytes Using SSC as a Proxy for Adipocyte Size (A and B) FSC/SSC plots (A) showing the gates used to sort out SSC_high (small) and SSC_low (large) adipocytes from the same sample, as well as representative confocal images (B) of the sorted cells showing the cell outline stained with FITC-lectin (white). Scale bars, 20 μm. (C) Measured adipocyte diameters of cells sorted from the total scWAT sample (all cells) or sorted using the depicted SSC_high and SSC_low gates. Note that the SSC-based gates efficiently enrich for the occurrence of small or large cells, which are less common in the total sorted sample. Lines show means ± SDs. ∗∗∗∗ p
    Figure Legend Snippet: Differential Sorting of Large and Small Adipocytes Using SSC as a Proxy for Adipocyte Size (A and B) FSC/SSC plots (A) showing the gates used to sort out SSC_high (small) and SSC_low (large) adipocytes from the same sample, as well as representative confocal images (B) of the sorted cells showing the cell outline stained with FITC-lectin (white). Scale bars, 20 μm. (C) Measured adipocyte diameters of cells sorted from the total scWAT sample (all cells) or sorted using the depicted SSC_high and SSC_low gates. Note that the SSC-based gates efficiently enrich for the occurrence of small or large cells, which are less common in the total sorted sample. Lines show means ± SDs. ∗∗∗∗ p

    Techniques Used: Staining

    Developing the Adipocyte Flow Cytometry Protocol (A) Freshly isolated adipocytes and SVF cells from AdTomato and WT mice, stained with fluorescein isothiocyanate (FITC)-labeled lectin (green) and Hoechst (blue) and imaged for tdTomato expression (red). Scale bars, 100 μm; note the differences in length. (B) Flow cytometry using standard settings for scWAT adipocytes and samples containing only pure mouse lipid or sunflower seed oil. Adipocyte samples show ungated and tdTomato + (T + ) gated events. (C) Flow cytometry using optimized adipocyte flow cytometry settings for scWAT adipocytes, BAT adipocytes, or SVF cells, showing ungated and tdTomato + (T + ) or Hoechst + (H + ) gated events. All of the experiments were repeated at least four times. See also Figures S1 and S2 .
    Figure Legend Snippet: Developing the Adipocyte Flow Cytometry Protocol (A) Freshly isolated adipocytes and SVF cells from AdTomato and WT mice, stained with fluorescein isothiocyanate (FITC)-labeled lectin (green) and Hoechst (blue) and imaged for tdTomato expression (red). Scale bars, 100 μm; note the differences in length. (B) Flow cytometry using standard settings for scWAT adipocytes and samples containing only pure mouse lipid or sunflower seed oil. Adipocyte samples show ungated and tdTomato + (T + ) gated events. (C) Flow cytometry using optimized adipocyte flow cytometry settings for scWAT adipocytes, BAT adipocytes, or SVF cells, showing ungated and tdTomato + (T + ) or Hoechst + (H + ) gated events. All of the experiments were repeated at least four times. See also Figures S1 and S2 .

    Techniques Used: Flow Cytometry, Cytometry, Isolation, Mouse Assay, Staining, Labeling, Expressing

    16) Product Images from "Compensatory endocytosis in bladder umbrella cells occurs through an integrin-regulated and RhoA- and dynamin-dependent pathway"

    Article Title: Compensatory endocytosis in bladder umbrella cells occurs through an integrin-regulated and RhoA- and dynamin-dependent pathway

    Journal: The EMBO Journal

    doi: 10.1038/emboj.2010.91

    Delivery of endocytosed membrane to late endosomes/lysosomes. ( A ) Co-localization of EEA1, Rab11a, ZO-1, giantin, or LAMP2 with FITC-WGA 10 min after experimental voiding. ( B ) Co-localization coefficients for FITC-WGA and the indicated marker 10 or 90 min after experimental voiding. Values are mean±s.e.m. ( n ⩾7). Those values significantly different from the 10-min time point ( P
    Figure Legend Snippet: Delivery of endocytosed membrane to late endosomes/lysosomes. ( A ) Co-localization of EEA1, Rab11a, ZO-1, giantin, or LAMP2 with FITC-WGA 10 min after experimental voiding. ( B ) Co-localization coefficients for FITC-WGA and the indicated marker 10 or 90 min after experimental voiding. Values are mean±s.e.m. ( n ⩾7). Those values significantly different from the 10-min time point ( P

    Techniques Used: Whole Genome Amplification, Marker

    17) Product Images from "Antigen presenting cells in the skin of a patient with hair loss and systemic lupus erythematosus"

    Article Title: Antigen presenting cells in the skin of a patient with hair loss and systemic lupus erythematosus

    Journal: North American Journal of Medical Sciences

    doi:

    a , DIF (EX 395-410/490-505/560-585) shows positive deposits of anti-human C3 FITC- conjugated at the BMZ of the skin(black arrow), (yellowish staining). In the same figure, slightly under the BMZ, ICAM-1/CD54 was also overexpressed (orange-reddish staining) in the dermal vessels (yellow arrow). The nuclei were counterstained with Hoechst 33258, (grayish stain) (200×). b , shows strong deposits of anti-human fibrinogen antibody, conjugated with FITC in the superficial and deep vessels of the skin (yellow arrows), and in c , anti-human fibrinogen was also visualized, but in this case when using pacific blue as secondary antibody (++++) (blue stain) (yellow arrows), (200×). d . H E demonstrates a mild epidermal hyperkeratosis with minimal follicular plugging. A mild, interface infiltrate of lymphocytes and histiocytes is noted, (black arrows) (100×). e ., DIF shows positive deposits of anti-human IgE-FITC conjugated (++++) (green staining) (yellow arrow) at the BMZ, with the nuclei were counterstained with Hoechst 33258, (grayish) (red arrow) (200×). Similar to e , but in this case, the nuclei were not counterstained 400×, (red arrow). g , H E stain shows some atrophy of the epidermis, liquefaction of the BMZ and homogenization of the papillary dermis near the BMZ (black arrow) (200×). h . DIF shows strong deposits of anti-human IgG FITC-conjugated positive at the basal membrane area of the sebaceous gland (400×), (red arrow). i , DIF shows positive deposits of anti-human IgE FITC- conjugated (++++), around the BMZ of the sebaceous glands (yellowish staining) (white arrows). In the same figure, slightly under the BMZ, ICAM-1/CD54 was also very positive (orange-reddish staining) was overexpressed in some dermal vessels (yellow arrows). The nuclei were counterstained in this case with DAPI (blue stain) (200×). j , similar to the i , but at higher magnification for better detail (400×). k . H E shows some mild spongiosis and BMZ degeneration of the hair follicle, with a mild cellular infiltrate of mainly lymphocytes and histiocytes (black arrow) (200×). l , DIF shows anti-human IgG-FITC conjugated (yellow staining) around the BMZ of the sebaceous gland (white arrows).
    Figure Legend Snippet: a , DIF (EX 395-410/490-505/560-585) shows positive deposits of anti-human C3 FITC- conjugated at the BMZ of the skin(black arrow), (yellowish staining). In the same figure, slightly under the BMZ, ICAM-1/CD54 was also overexpressed (orange-reddish staining) in the dermal vessels (yellow arrow). The nuclei were counterstained with Hoechst 33258, (grayish stain) (200×). b , shows strong deposits of anti-human fibrinogen antibody, conjugated with FITC in the superficial and deep vessels of the skin (yellow arrows), and in c , anti-human fibrinogen was also visualized, but in this case when using pacific blue as secondary antibody (++++) (blue stain) (yellow arrows), (200×). d . H E demonstrates a mild epidermal hyperkeratosis with minimal follicular plugging. A mild, interface infiltrate of lymphocytes and histiocytes is noted, (black arrows) (100×). e ., DIF shows positive deposits of anti-human IgE-FITC conjugated (++++) (green staining) (yellow arrow) at the BMZ, with the nuclei were counterstained with Hoechst 33258, (grayish) (red arrow) (200×). Similar to e , but in this case, the nuclei were not counterstained 400×, (red arrow). g , H E stain shows some atrophy of the epidermis, liquefaction of the BMZ and homogenization of the papillary dermis near the BMZ (black arrow) (200×). h . DIF shows strong deposits of anti-human IgG FITC-conjugated positive at the basal membrane area of the sebaceous gland (400×), (red arrow). i , DIF shows positive deposits of anti-human IgE FITC- conjugated (++++), around the BMZ of the sebaceous glands (yellowish staining) (white arrows). In the same figure, slightly under the BMZ, ICAM-1/CD54 was also very positive (orange-reddish staining) was overexpressed in some dermal vessels (yellow arrows). The nuclei were counterstained in this case with DAPI (blue stain) (200×). j , similar to the i , but at higher magnification for better detail (400×). k . H E shows some mild spongiosis and BMZ degeneration of the hair follicle, with a mild cellular infiltrate of mainly lymphocytes and histiocytes (black arrow) (200×). l , DIF shows anti-human IgG-FITC conjugated (yellow staining) around the BMZ of the sebaceous gland (white arrows).

    Techniques Used: Staining, Homogenization

    18) Product Images from "Mechanical force-driven TNFα endocytosis governs stem cell homeostasis"

    Article Title: Mechanical force-driven TNFα endocytosis governs stem cell homeostasis

    Journal: Bone Research

    doi: 10.1038/s41413-020-00117-x

    Mechanical force drives TNFα endocytosis and promotes MSC function. a , b Examination of secreted concentrations and protein expression levels of TNFα. MSCs underwent cyclic loading of stretch force at 15% elongation, 0.5 Hz. N = 3. c Western blot analyses of mTOR signaling ( N = 3). d – f Functional analyses of MSCs according to BrdU labeling and osteogenic and adipogenic differentiation. N = 3. Scale bars = 100 μm. Stretch force was applied twice per week for 2 h per session. g qRT-PCR analysis of the mRNA expression levels of TNFα ( N = 3). h , i Endocytosis analysis of FITC-labeled TNFα uptake by MSCs for 2 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . Stretch force was applied for a 2 h period. N = 3. Scale bars = 20 μm. j , k Endocytosis analysis of Alexa Fluor TM 488-conjugated bovine serum albumin (488-BSA) taken up by MSCs for 2 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . Stretch force was applied for a 2 h period. N = 3. Scale bars = 20 μm. l Diagram showing mechanical force-driven TNFα endocytosis. For quantification of Western blotting, a two-tailed Student’s t test was used for the comparison between the treatment and stretch 0 h/control groups. * P
    Figure Legend Snippet: Mechanical force drives TNFα endocytosis and promotes MSC function. a , b Examination of secreted concentrations and protein expression levels of TNFα. MSCs underwent cyclic loading of stretch force at 15% elongation, 0.5 Hz. N = 3. c Western blot analyses of mTOR signaling ( N = 3). d – f Functional analyses of MSCs according to BrdU labeling and osteogenic and adipogenic differentiation. N = 3. Scale bars = 100 μm. Stretch force was applied twice per week for 2 h per session. g qRT-PCR analysis of the mRNA expression levels of TNFα ( N = 3). h , i Endocytosis analysis of FITC-labeled TNFα uptake by MSCs for 2 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . Stretch force was applied for a 2 h period. N = 3. Scale bars = 20 μm. j , k Endocytosis analysis of Alexa Fluor TM 488-conjugated bovine serum albumin (488-BSA) taken up by MSCs for 2 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . Stretch force was applied for a 2 h period. N = 3. Scale bars = 20 μm. l Diagram showing mechanical force-driven TNFα endocytosis. For quantification of Western blotting, a two-tailed Student’s t test was used for the comparison between the treatment and stretch 0 h/control groups. * P

    Techniques Used: Expressing, Western Blot, Functional Assay, Labeling, Quantitative RT-PCR, In Vitro, Two Tailed Test

    TNFα safeguards MSC homeostasis in a receptor-independent manner through endocytosis. a – d Functional analyses of MSCs according to CFU, BrdU labeling, and osteogenic and adipogenic differentiation. MSCs were derived from WT, TNFα −/− or TNFR −/− mice ( N = 5). Scale bars = 100 μm. e Calcein labeling for bone formation analysis in WT and TNFR −/− mice ( N = 5). Scale bar = 50 μm. f Oli red O staining for bone marrow adiposity in WT and TNFR −/− mice ( N = 5). Scale bar = 150 μm. g Endocytosis analysis of FITC-labeled TNFα uptake by MSCs for 24 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . N = 3. Scale bars = 20 μm (top) and 7 μm (bottom). h Western blot analysis of protein expression levels ( N = 3). i – k Functional analyses of MSCs according to BrdU labeling and osteogenic and adipogenic differentiation. N = 3. Scale bars = 100 μm. l Diagram showing that TNFα regulates MSC homeostasis in a receptor-independent manner through endocytosis. For quantification of Western blotting, a two-tailed Student’s t test was used for the comparison between the treatment and WT groups. * P
    Figure Legend Snippet: TNFα safeguards MSC homeostasis in a receptor-independent manner through endocytosis. a – d Functional analyses of MSCs according to CFU, BrdU labeling, and osteogenic and adipogenic differentiation. MSCs were derived from WT, TNFα −/− or TNFR −/− mice ( N = 5). Scale bars = 100 μm. e Calcein labeling for bone formation analysis in WT and TNFR −/− mice ( N = 5). Scale bar = 50 μm. f Oli red O staining for bone marrow adiposity in WT and TNFR −/− mice ( N = 5). Scale bar = 150 μm. g Endocytosis analysis of FITC-labeled TNFα uptake by MSCs for 24 h in vitro. Pitstop ® 2 was used to inhibit clathrin-mediated endocytosis at 12 μmol·L −1 . N = 3. Scale bars = 20 μm (top) and 7 μm (bottom). h Western blot analysis of protein expression levels ( N = 3). i – k Functional analyses of MSCs according to BrdU labeling and osteogenic and adipogenic differentiation. N = 3. Scale bars = 100 μm. l Diagram showing that TNFα regulates MSC homeostasis in a receptor-independent manner through endocytosis. For quantification of Western blotting, a two-tailed Student’s t test was used for the comparison between the treatment and WT groups. * P

    Techniques Used: Functional Assay, Labeling, Derivative Assay, Mouse Assay, Staining, In Vitro, Western Blot, Expressing, Two Tailed Test

    19) Product Images from "Remodeling of the Extracellular Matrix by Endothelial Cell-Targeting siRNA Improves the EPR-Based Delivery of 100 nm Particles"

    Article Title: Remodeling of the Extracellular Matrix by Endothelial Cell-Targeting siRNA Improves the EPR-Based Delivery of 100 nm Particles

    Journal: Molecular Therapy

    doi: 10.1038/mt.2016.178

    Involvement of macrophages on the improvement in the intratumoral distribution of LNPs. ( a ) Increase in macrophages by si-VR2 encapsulated in the RGD-MEND. Blue, green and red dots indicate nuclei (Hoechst33342), vessel (FITC-isolectin) and macrophages (F4/80). Scale bars are 100 μm. ( b ) Pixels indicating macrophages from a were counted by Image J. ANOVA was performed for statistical analysis, followed by the SNK test ( n = 9–12). ** P
    Figure Legend Snippet: Involvement of macrophages on the improvement in the intratumoral distribution of LNPs. ( a ) Increase in macrophages by si-VR2 encapsulated in the RGD-MEND. Blue, green and red dots indicate nuclei (Hoechst33342), vessel (FITC-isolectin) and macrophages (F4/80). Scale bars are 100 μm. ( b ) Pixels indicating macrophages from a were counted by Image J. ANOVA was performed for statistical analysis, followed by the SNK test ( n = 9–12). ** P

    Techniques Used:

    Vascular maturation by siRNA against VEGFR2 encapsulated in the RGD-MEND. ( a ) Representative image of the increase in pericyte coverage by the RGD-MEND. Tumor tissues were cryo-sectioned after the continuous inhibition of VEGFR2. The sections were stained with Hoechst33342 (blue, nucleus), FITC-isolectin (green, vessels) and cy3-αSMA (red, pericytes). Scale bars: 100 μm. ( b ) Quantitative data of a . Pixels were counted in nine images from three independent mice, and the red pixels (pericytes) were then normalized to green pixels (vessels). ( c ) Dose-dependency for the increase in pericyte coverage. Pericytes were counted when the dosages of si-VR2 varied from 0.75 mg/kg to 3.0 mg/kg (each groups were of three mice). ( d ) Decrease in hypoxic area in RGD-MEND-treated mice. Tumor tissues were collected 90 minutes after the injection of the hypoxia-probe pimonidazole. Green and red pixels vessels and the indicated hypoxic regions, respectively. Scale bars: 100 μm. ( e ) Red dots indicating hypoxic regions were counted and normalized to nucleus areas. Data were obtained from nine images from three independent mice. ( f ) Recovery of blood flow by the RGD-MEND. FITC-isolectin B4 was systemically injected before sacrifice, and the collected tumor tissues were then immersed in Alexa647-isolectin B4. Arrows show the vasculature without blood flow. ( g ) Quantitative data of perfused vessels. Population of the vasculature with blood flows (shown as yellow) against all of the vasculature (shown as yellow and red) were counted. FITC, fluorescein isothiocyanate; RGD-MEND, RGD-modified liposomal siRNA; VEGFR2, VEGF receptor 2.
    Figure Legend Snippet: Vascular maturation by siRNA against VEGFR2 encapsulated in the RGD-MEND. ( a ) Representative image of the increase in pericyte coverage by the RGD-MEND. Tumor tissues were cryo-sectioned after the continuous inhibition of VEGFR2. The sections were stained with Hoechst33342 (blue, nucleus), FITC-isolectin (green, vessels) and cy3-αSMA (red, pericytes). Scale bars: 100 μm. ( b ) Quantitative data of a . Pixels were counted in nine images from three independent mice, and the red pixels (pericytes) were then normalized to green pixels (vessels). ( c ) Dose-dependency for the increase in pericyte coverage. Pericytes were counted when the dosages of si-VR2 varied from 0.75 mg/kg to 3.0 mg/kg (each groups were of three mice). ( d ) Decrease in hypoxic area in RGD-MEND-treated mice. Tumor tissues were collected 90 minutes after the injection of the hypoxia-probe pimonidazole. Green and red pixels vessels and the indicated hypoxic regions, respectively. Scale bars: 100 μm. ( e ) Red dots indicating hypoxic regions were counted and normalized to nucleus areas. Data were obtained from nine images from three independent mice. ( f ) Recovery of blood flow by the RGD-MEND. FITC-isolectin B4 was systemically injected before sacrifice, and the collected tumor tissues were then immersed in Alexa647-isolectin B4. Arrows show the vasculature without blood flow. ( g ) Quantitative data of perfused vessels. Population of the vasculature with blood flows (shown as yellow) against all of the vasculature (shown as yellow and red) were counted. FITC, fluorescein isothiocyanate; RGD-MEND, RGD-modified liposomal siRNA; VEGFR2, VEGF receptor 2.

    Techniques Used: Inhibition, Staining, Mouse Assay, Injection, Flow Cytometry, Modification

    20) Product Images from "Csf1r-mApple Transgene Expression and Ligand Binding In Vivo Reveal Dynamics of CSF1R Expression within the Mononuclear Phagocyte System"

    Article Title: Csf1r-mApple Transgene Expression and Ligand Binding In Vivo Reveal Dynamics of CSF1R Expression within the Mononuclear Phagocyte System

    Journal: The Journal of Immunology Author Choice

    doi: 10.4049/jimmunol.1701488

    Csf1r -mApple and Δ Csf1r -ECFP transgenes allow imaging of distinct lineages of hepatic myeloid cells. Confocal image of the surface of the left lobe of the liver of a Δ Csf1r -ECFP ( A ), Csf1r -mApple ( B ), WT ( C ), and Csf1r- mApple/Δ Csf1r -ECFP ( D ) mouse imaged ex vivo. FITC-Lectin I was injected i.v. to reveal liver sinusoidal endothelium. Scale bars represent 20 μm (A–C) or 50 μm (D).
    Figure Legend Snippet: Csf1r -mApple and Δ Csf1r -ECFP transgenes allow imaging of distinct lineages of hepatic myeloid cells. Confocal image of the surface of the left lobe of the liver of a Δ Csf1r -ECFP ( A ), Csf1r -mApple ( B ), WT ( C ), and Csf1r- mApple/Δ Csf1r -ECFP ( D ) mouse imaged ex vivo. FITC-Lectin I was injected i.v. to reveal liver sinusoidal endothelium. Scale bars represent 20 μm (A–C) or 50 μm (D).

    Techniques Used: Imaging, Ex Vivo, Injection

    Csf1r- mApple and Δ Csf1r -ECFP transgenes allow imaging of distinct lineages of pulmonary myeloid cells. Confocal image of a transverse section of lung from a Δ Csf1r -ECFP ( A ), Csf1r -mApple ( B ), WT ( C ), and Csf1r -mApple/Δ Csf1r -ECFP ( D ) mouse imaged ex vivo. FITC-Lectin was injected i.v. to reveal pulmonary vasculature. Scale bars in all panels represent 50 μm.
    Figure Legend Snippet: Csf1r- mApple and Δ Csf1r -ECFP transgenes allow imaging of distinct lineages of pulmonary myeloid cells. Confocal image of a transverse section of lung from a Δ Csf1r -ECFP ( A ), Csf1r -mApple ( B ), WT ( C ), and Csf1r -mApple/Δ Csf1r -ECFP ( D ) mouse imaged ex vivo. FITC-Lectin was injected i.v. to reveal pulmonary vasculature. Scale bars in all panels represent 50 μm.

    Techniques Used: Imaging, Ex Vivo, Injection

    21) Product Images from "CB1 modulation of temporally distinct synaptic facilitation among local circuit interneurons mediated by N-type calcium channels in CA1"

    Article Title: CB1 modulation of temporally distinct synaptic facilitation among local circuit interneurons mediated by N-type calcium channels in CA1

    Journal: Journal of Neurophysiology

    doi: 10.1152/jn.00831.2010

    Temporally distinct synaptic facilitation that is delayed on the onset of release. A : reconstruction of a synaptically connected stratum lacunosum moleculare (SLM) interneuron, lacunosum moleculare-Shaffer collateral associated cell (LM-SCA; blue-soma, light blue-axon), and a stratum radiatum (SR), Shaffer collateral associated cell (SCA; deep red- dendrites, red-axon). B : double immunoflorescence labeling revealed the two recorded cells filled with biocytin (AMCA) immunopositive for CCK (FITC) and immunonegative for VIP (Texas red). C : intrinsic membrane properties of the presynaptic LM-SCA cell. In response to hyperpolarizing current injection this interneuron displayed pronounced sag, indicative of I h current. In response to depolarizing current fast but adapting firing patterns were displayed. D : IPSPs elicited by LM-SCA cells displayed a delayed onset of synaptic facilitation (DORF). IPSPs were usually observed after third to fifth presynaptic spike in a train. E : another synaptic connection between a LM-SCA cell and SCA cell; the onset of first IPSP did not appear earlier in the train when increasing the firing frequency of the presynaptic interneuron from 25 Hz to 50 Hz. At higher presynaptic firing frequencies the IPSPs summated more readily, shown by the black trace. F : comparison of average failure rates of 1st, 2nd, 3rd and 4th IPSPs in control at DORF, frequency dependent facilitating (FDF), and depressing synapses studied. SP, stratum pyramidale, SO, stratum oriens.
    Figure Legend Snippet: Temporally distinct synaptic facilitation that is delayed on the onset of release. A : reconstruction of a synaptically connected stratum lacunosum moleculare (SLM) interneuron, lacunosum moleculare-Shaffer collateral associated cell (LM-SCA; blue-soma, light blue-axon), and a stratum radiatum (SR), Shaffer collateral associated cell (SCA; deep red- dendrites, red-axon). B : double immunoflorescence labeling revealed the two recorded cells filled with biocytin (AMCA) immunopositive for CCK (FITC) and immunonegative for VIP (Texas red). C : intrinsic membrane properties of the presynaptic LM-SCA cell. In response to hyperpolarizing current injection this interneuron displayed pronounced sag, indicative of I h current. In response to depolarizing current fast but adapting firing patterns were displayed. D : IPSPs elicited by LM-SCA cells displayed a delayed onset of synaptic facilitation (DORF). IPSPs were usually observed after third to fifth presynaptic spike in a train. E : another synaptic connection between a LM-SCA cell and SCA cell; the onset of first IPSP did not appear earlier in the train when increasing the firing frequency of the presynaptic interneuron from 25 Hz to 50 Hz. At higher presynaptic firing frequencies the IPSPs summated more readily, shown by the black trace. F : comparison of average failure rates of 1st, 2nd, 3rd and 4th IPSPs in control at DORF, frequency dependent facilitating (FDF), and depressing synapses studied. SP, stratum pyramidale, SO, stratum oriens.

    Techniques Used: Labeling, Injection

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Vector Laboratories ulex europaeus aggluitinin 1 uea 1 conjugated to fitc
    Commensals utilize fucose detached from proteins fucosylated by Fut2 upon systemic challenge with LPS a, Silver-stained SDS-PAGE of <t>UEA-1-precipitated</t> SI IEC protein from control or LPS-treated mouse. b, Ratios of digestive enzymes activities in SI IECs of LPS-treated to untreated Fut2-sufficient (black bars) or Fut2-deficient (open bars) mice two days after LPS injection. Mean±s.e.m. of 4 combined experiments, 4 mice/group. c, SDS-PAGE of intestinal contents blotted on nitrocellulose and stained with UEA-1-peroxidase complexes. s, SI; c, cecum; f, feces; abx, antibiotic treated mice; DFJ, deoxyfuconojirimycin. d, e, Fucose-sensitive GFP reporter expression in gnotobiotic mice colonized with the indicated strains (d, ) or SPF mice (e). Dots are values for individual bacteria, lines are means, n=120; * P
    Ulex Europaeus Aggluitinin 1 Uea 1 Conjugated To Fitc, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ulex europaeus aggluitinin 1 uea 1 conjugated to fitc/product/Vector Laboratories
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    ulex europaeus aggluitinin 1 uea 1 conjugated to fitc - by Bioz Stars, 2022-10
    90/100 stars
      Buy from Supplier

    95
    Vector Laboratories fitc labeled ulex europaeus lectin
    Inhibition of miR-15a decreases endothelial cell proliferation and enhances cell death. ( A ) HUVECs were transfected with either a control negative inhibitor or a miR-15a inhibitor. 48 h later proliferation ( A ) or cell death ( B ) was measured using a luciferase-based Cell Titer glo assay ( A ) or Caspase 3 7 CasGlo assay ( B ). ( C ) Fibrin bead 3D angiogenesis assay. HUVECs were transfected as described in A and were coated onto cytodex beads and allowed to sprout in a fibrin gel in the presence of smooth muscle cells over 5 days. The images show representative beads with angiogenic sprouts stained with Ulex <t>europaeus</t> <t>lectin</t> for each condition. Bars depict mean +SEM of lectin area analyzed across at least 25 beads per group. Scale bar = 100 μm. D) HUVECs were transfected as described in A with the indicated concentrations of either control inhibitor or miR-15a inhibitor. 24 h later, cells were irradiated at the indicated doses. 48 h post irradiation, proliferation was measured using a Cell Titer glo assay. Synergy was calculated using the Chou-Talalay method with combination index
    Fitc Labeled Ulex Europaeus Lectin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 95/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fitc labeled ulex europaeus lectin/product/Vector Laboratories
    Average 95 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    fitc labeled ulex europaeus lectin - by Bioz Stars, 2022-10
    95/100 stars
      Buy from Supplier

    96
    Vector Laboratories fitc conjugated sambucus nigra sna lectin
    Mass spectrometry analysis of <t>SNA-binding</t> proteins isolated from DCs reveals key immune-related proteins. ( A ) The reactivity of <t>Sambucus</t> <t>nigra</t> (SNA) <t>lectin</t> to α2,6-sialylated glycans on the surface of human DCs was quantified by flow cytometry after different maturation stimuli: IFN-γ, IL-1β, TNF-α and LPS. Unstimulated DCs were used as control. Values presented are mean ± SEM (N = 6). Statistically significant differences are indicated by asterisks (* p ≤ 0.05). ( B ) Schematic representation of the steps followed to identify α2,6-sialylated proteins from DCs. Whole cell lysates of human DCs were immunoprecipitated through a SNA-binding column. The eluted proteins were analyzed by mass spectrometry and the corresponding identified scores were matched and associated with Gene Ontology (GO) entries. ( C ) Distribution of the identified sialylated proteins by their molecular function. Pie chart represents different molecular functions of the identified proteins, according to the GO entries. Immune system processes were highlighted.
    Fitc Conjugated Sambucus Nigra Sna Lectin, supplied by Vector Laboratories, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fitc conjugated sambucus nigra sna lectin/product/Vector Laboratories
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fitc conjugated sambucus nigra sna lectin - by Bioz Stars, 2022-10
    96/100 stars
      Buy from Supplier

    Image Search Results


    Commensals utilize fucose detached from proteins fucosylated by Fut2 upon systemic challenge with LPS a, Silver-stained SDS-PAGE of UEA-1-precipitated SI IEC protein from control or LPS-treated mouse. b, Ratios of digestive enzymes activities in SI IECs of LPS-treated to untreated Fut2-sufficient (black bars) or Fut2-deficient (open bars) mice two days after LPS injection. Mean±s.e.m. of 4 combined experiments, 4 mice/group. c, SDS-PAGE of intestinal contents blotted on nitrocellulose and stained with UEA-1-peroxidase complexes. s, SI; c, cecum; f, feces; abx, antibiotic treated mice; DFJ, deoxyfuconojirimycin. d, e, Fucose-sensitive GFP reporter expression in gnotobiotic mice colonized with the indicated strains (d, ) or SPF mice (e). Dots are values for individual bacteria, lines are means, n=120; * P

    Journal: Nature

    Article Title: Rapid fucosylation of intestinal epithelium sustains host-commensal symbiosis in sickness

    doi: 10.1038/nature13823

    Figure Lengend Snippet: Commensals utilize fucose detached from proteins fucosylated by Fut2 upon systemic challenge with LPS a, Silver-stained SDS-PAGE of UEA-1-precipitated SI IEC protein from control or LPS-treated mouse. b, Ratios of digestive enzymes activities in SI IECs of LPS-treated to untreated Fut2-sufficient (black bars) or Fut2-deficient (open bars) mice two days after LPS injection. Mean±s.e.m. of 4 combined experiments, 4 mice/group. c, SDS-PAGE of intestinal contents blotted on nitrocellulose and stained with UEA-1-peroxidase complexes. s, SI; c, cecum; f, feces; abx, antibiotic treated mice; DFJ, deoxyfuconojirimycin. d, e, Fucose-sensitive GFP reporter expression in gnotobiotic mice colonized with the indicated strains (d, ) or SPF mice (e). Dots are values for individual bacteria, lines are means, n=120; * P

    Article Snippet: Lectin staining For whole-mount staining, small intestine was removed, a 1 cm piece from the upper third was excised, opened, cleaned of mucus in cold PBS, and incubated with Ulex europaeus aggluitinin-1 (UEA-1) conjugated to FITC, TRITC or atto-594 (Vector Laboratories, Burlingame, CA or Sigma-Aldrich) for 15 min on ice.

    Techniques: Staining, SDS Page, Mouse Assay, Injection, Expressing

    Requirements and kinetics for SI fucosylation induced by systemic injection of TLR ligands a, Systemic injection of bacterial TLR ligands induces small intestine fucosylation, but simple starvation does not. UEA-1 staining (as in Fig. 1 ) after i.p. injection of CpG DNA, or Pam 3 CSK 4 , or food deprivation for 24 h of BALB/c SPF mouse. b, LPS injection causes SI fucosylation in various inbred mouse strains. SPF mice of the indicated strains were injected with LPS i.p. and SI was stained with UEA-1 after 24 hours, as in Fig. 1 . Representative of at least 2 independent experiments. c, Fucosylation peaks at 8 h after LPS injection and is still detectable at 96 h. d, M cells can be readily detected by SEM and UEA-1 staining of the domes of the Peyer's patches, but are rare in the villi and are not massively induced in the villi by LPS injection. UEA-1 staining and SEM were performed on adjacent pieces from proximal 1/3 of SI. Scale bars=100 μm for UEA-1 staining, 50 μm for SEM images. Representative of at least 2 independent experiments. e , SI fucosylation does not require the presence of endogenous microbiota (LPS injection in GF mouse) and is not induced by oral administration of LPS (1 mg).

    Journal: Nature

    Article Title: Rapid fucosylation of intestinal epithelium sustains host-commensal symbiosis in sickness

    doi: 10.1038/nature13823

    Figure Lengend Snippet: Requirements and kinetics for SI fucosylation induced by systemic injection of TLR ligands a, Systemic injection of bacterial TLR ligands induces small intestine fucosylation, but simple starvation does not. UEA-1 staining (as in Fig. 1 ) after i.p. injection of CpG DNA, or Pam 3 CSK 4 , or food deprivation for 24 h of BALB/c SPF mouse. b, LPS injection causes SI fucosylation in various inbred mouse strains. SPF mice of the indicated strains were injected with LPS i.p. and SI was stained with UEA-1 after 24 hours, as in Fig. 1 . Representative of at least 2 independent experiments. c, Fucosylation peaks at 8 h after LPS injection and is still detectable at 96 h. d, M cells can be readily detected by SEM and UEA-1 staining of the domes of the Peyer's patches, but are rare in the villi and are not massively induced in the villi by LPS injection. UEA-1 staining and SEM were performed on adjacent pieces from proximal 1/3 of SI. Scale bars=100 μm for UEA-1 staining, 50 μm for SEM images. Representative of at least 2 independent experiments. e , SI fucosylation does not require the presence of endogenous microbiota (LPS injection in GF mouse) and is not induced by oral administration of LPS (1 mg).

    Article Snippet: Lectin staining For whole-mount staining, small intestine was removed, a 1 cm piece from the upper third was excised, opened, cleaned of mucus in cold PBS, and incubated with Ulex europaeus aggluitinin-1 (UEA-1) conjugated to FITC, TRITC or atto-594 (Vector Laboratories, Burlingame, CA or Sigma-Aldrich) for 15 min on ice.

    Techniques: Injection, Staining, Mouse Assay

    Fucosylated protein in IECs and gut contents a , Proteins α(1,2)fucosylated in IECs after LPS injection identified by UEA-1 precipitation and mass spectrometry. Abundance is the number of peptide fragments attributed to each gene. b, IECs from Fut2 + untreated, Fut2 + LPS-treated, or Fut2 − untreated mice were isolated, and lysates separated by SDS-PAGE. α(1,2)fucosylated proteins were detected by blotting with UEA-1 lectin conjugated to HRP. c, Identical gel stained with Coomassie blue for total protein content. d, Relative density of the boxed area of each lane from b divided by the relative density in c. e, UEA-1 staining of luminal proteins as in Fig. 3c . Blot is overexposed to show absence of luminal fucosylated proteins in the LPS-treated, Fut2 − mouse.

    Journal: Nature

    Article Title: Rapid fucosylation of intestinal epithelium sustains host-commensal symbiosis in sickness

    doi: 10.1038/nature13823

    Figure Lengend Snippet: Fucosylated protein in IECs and gut contents a , Proteins α(1,2)fucosylated in IECs after LPS injection identified by UEA-1 precipitation and mass spectrometry. Abundance is the number of peptide fragments attributed to each gene. b, IECs from Fut2 + untreated, Fut2 + LPS-treated, or Fut2 − untreated mice were isolated, and lysates separated by SDS-PAGE. α(1,2)fucosylated proteins were detected by blotting with UEA-1 lectin conjugated to HRP. c, Identical gel stained with Coomassie blue for total protein content. d, Relative density of the boxed area of each lane from b divided by the relative density in c. e, UEA-1 staining of luminal proteins as in Fig. 3c . Blot is overexposed to show absence of luminal fucosylated proteins in the LPS-treated, Fut2 − mouse.

    Article Snippet: Lectin staining For whole-mount staining, small intestine was removed, a 1 cm piece from the upper third was excised, opened, cleaned of mucus in cold PBS, and incubated with Ulex europaeus aggluitinin-1 (UEA-1) conjugated to FITC, TRITC or atto-594 (Vector Laboratories, Burlingame, CA or Sigma-Aldrich) for 15 min on ice.

    Techniques: Injection, Mass Spectrometry, Mouse Assay, Isolation, SDS Page, Staining

    Inhibition of miR-15a decreases endothelial cell proliferation and enhances cell death. ( A ) HUVECs were transfected with either a control negative inhibitor or a miR-15a inhibitor. 48 h later proliferation ( A ) or cell death ( B ) was measured using a luciferase-based Cell Titer glo assay ( A ) or Caspase 3 7 CasGlo assay ( B ). ( C ) Fibrin bead 3D angiogenesis assay. HUVECs were transfected as described in A and were coated onto cytodex beads and allowed to sprout in a fibrin gel in the presence of smooth muscle cells over 5 days. The images show representative beads with angiogenic sprouts stained with Ulex europaeus lectin for each condition. Bars depict mean +SEM of lectin area analyzed across at least 25 beads per group. Scale bar = 100 μm. D) HUVECs were transfected as described in A with the indicated concentrations of either control inhibitor or miR-15a inhibitor. 24 h later, cells were irradiated at the indicated doses. 48 h post irradiation, proliferation was measured using a Cell Titer glo assay. Synergy was calculated using the Chou-Talalay method with combination index

    Journal: Scientific Reports

    Article Title: Differential regulation of microRNA-15a by radiation affects angiogenesis and tumor growth via modulation of acid sphingomyelinase

    doi: 10.1038/s41598-020-62621-8

    Figure Lengend Snippet: Inhibition of miR-15a decreases endothelial cell proliferation and enhances cell death. ( A ) HUVECs were transfected with either a control negative inhibitor or a miR-15a inhibitor. 48 h later proliferation ( A ) or cell death ( B ) was measured using a luciferase-based Cell Titer glo assay ( A ) or Caspase 3 7 CasGlo assay ( B ). ( C ) Fibrin bead 3D angiogenesis assay. HUVECs were transfected as described in A and were coated onto cytodex beads and allowed to sprout in a fibrin gel in the presence of smooth muscle cells over 5 days. The images show representative beads with angiogenic sprouts stained with Ulex europaeus lectin for each condition. Bars depict mean +SEM of lectin area analyzed across at least 25 beads per group. Scale bar = 100 μm. D) HUVECs were transfected as described in A with the indicated concentrations of either control inhibitor or miR-15a inhibitor. 24 h later, cells were irradiated at the indicated doses. 48 h post irradiation, proliferation was measured using a Cell Titer glo assay. Synergy was calculated using the Chou-Talalay method with combination index

    Article Snippet: Sprouts were visualized from days 3–4 via confocal imaging after overnight incubation with FITC labeled Ulex europaeus lectin (Vector labs).

    Techniques: Inhibition, Transfection, Luciferase, Glo Assay, Angiogenesis Assay, Staining, Irradiation

    Endothelial cell characteristics. After 7 days culture (A), cells taken up DiI-ac-LDL were red fluorescence (B), bound to FITC-UEA-I displayed green fluorescence (C), and the double-stained cells showed yellow fluorescence in cytoplasm (D). (Magnification ×200).

    Journal: PLoS ONE

    Article Title: In Vivo Serial MR Imaging of Magnetically Labeled Endothelial Progenitor Cells Homing to the Endothelium Injured Artery in Mice

    doi: 10.1371/journal.pone.0020790

    Figure Lengend Snippet: Endothelial cell characteristics. After 7 days culture (A), cells taken up DiI-ac-LDL were red fluorescence (B), bound to FITC-UEA-I displayed green fluorescence (C), and the double-stained cells showed yellow fluorescence in cytoplasm (D). (Magnification ×200).

    Article Snippet: The fixed cells were then incubated with 1 mL (10 µg/ml) FITC-UEA I (Vector Laboratories Inc., Burlingame, CA) at 37°C for 1 hour.

    Techniques: Fluorescence, Staining

    Mass spectrometry analysis of SNA-binding proteins isolated from DCs reveals key immune-related proteins. ( A ) The reactivity of Sambucus nigra (SNA) lectin to α2,6-sialylated glycans on the surface of human DCs was quantified by flow cytometry after different maturation stimuli: IFN-γ, IL-1β, TNF-α and LPS. Unstimulated DCs were used as control. Values presented are mean ± SEM (N = 6). Statistically significant differences are indicated by asterisks (* p ≤ 0.05). ( B ) Schematic representation of the steps followed to identify α2,6-sialylated proteins from DCs. Whole cell lysates of human DCs were immunoprecipitated through a SNA-binding column. The eluted proteins were analyzed by mass spectrometry and the corresponding identified scores were matched and associated with Gene Ontology (GO) entries. ( C ) Distribution of the identified sialylated proteins by their molecular function. Pie chart represents different molecular functions of the identified proteins, according to the GO entries. Immune system processes were highlighted.

    Journal: Pharmaceutics

    Article Title: MHC Class I Stability is Modulated by Cell Surface Sialylation in Human Dendritic Cells

    doi: 10.3390/pharmaceutics12030249

    Figure Lengend Snippet: Mass spectrometry analysis of SNA-binding proteins isolated from DCs reveals key immune-related proteins. ( A ) The reactivity of Sambucus nigra (SNA) lectin to α2,6-sialylated glycans on the surface of human DCs was quantified by flow cytometry after different maturation stimuli: IFN-γ, IL-1β, TNF-α and LPS. Unstimulated DCs were used as control. Values presented are mean ± SEM (N = 6). Statistically significant differences are indicated by asterisks (* p ≤ 0.05). ( B ) Schematic representation of the steps followed to identify α2,6-sialylated proteins from DCs. Whole cell lysates of human DCs were immunoprecipitated through a SNA-binding column. The eluted proteins were analyzed by mass spectrometry and the corresponding identified scores were matched and associated with Gene Ontology (GO) entries. ( C ) Distribution of the identified sialylated proteins by their molecular function. Pie chart represents different molecular functions of the identified proteins, according to the GO entries. Immune system processes were highlighted.

    Article Snippet: For flow cytometry analysis, we also stained cells with FITC-conjugated Sambucus nigra (SNA) lectin (Vector Labs) and the fluorescently FITC-labeled peptides NLVPKFITCVATV and SIINFEKFITCL (Genecust).

    Techniques: Mass Spectrometry, Binding Assay, Isolation, Flow Cytometry, Immunoprecipitation