Journal: Cell reports

Article Title: The nociceptive activity of peripheral sensory neurons is modulated by the neuronal membrane proteasome

doi: 10.1016/j.celrep.2024.114058

Figure Lengend Snippet: (A) Representative micrographs of immuno-EM labeling on primary DRG cultures. White arrowheads: gold particles localized on the cytoplasm. Black arrowheads: gold particles localized on the membrane. Bar graphs to the right show the proportion of gold particles localized on the cytoplasm (Cyto) and membrane (Mem). Violin plots show the distribution of gold particles within 240 nm of the neuronal membranes. Data are presented as mean ± SEM ( n = 14–20 micrographs analyzed from two independent animals). (B) Representative images of antibody feeding experiments in DRG cultures against the β5 proteasome subunit followed by cytoplasmic NF-H staining. White box: magnified section shown on bottom of composite image. (C) Representative 3D projection images of antibody feeding experiments in DRG cultures against the β5 proteasome subunit followed by cytoplasmic NF-H staining. (D) Representative 3D projection images of antibody feeding experiments in DRG cultures against the P2X3 and β5 proteasome subunit co-labeling. Bar graph shows quantification of surface P2X3 and β5 co-labeling under the conditions that at least one neuron in the field of view was positive for both NMP and P2X3. Data are presented as mean ± SEM ( n = 13 images analyzed). See also .

Article Snippet: Rabbit anti-β5 proteasome subunit , Enzo , BML-PW8895; RRID: AB_10540901.

Techniques: Labeling, Membrane, Staining

Journal: Cell reports

Article Title: The nociceptive activity of peripheral sensory neurons is modulated by the neuronal membrane proteasome

doi: 10.1016/j.celrep.2024.114058

Figure Lengend Snippet:

Article Snippet: Rabbit anti-β5 proteasome subunit , Enzo , BML-PW8895; RRID: AB_10540901.

Techniques: Recombinant, Saline, Protease Inhibitor, Synthesized, Bradford Protein Assay, Software, Imaging

Journal: Journal of Virology

Article Title: Human Adenovirus Type 37 Uses α V β 1 and α 3 β 1 Integrins for Infection of Human Corneal Cells

doi: 10.1128/JVI.02019-16

Figure Lengend Snippet: Cross sections of human corneas labeled (in green) with antibodies against integrin α2 (P1E6), α3 (ASC-1), α4 (P4C2), α5 (P1D6), α6 (MP4F10), αV (272-17E6), β1 (P5D2), β3 (MHF4), β4 (422325), and β5 (H00051706-D01P), as well as secondary-antibody-only controls GAM Alexa 488 and GAR Alexa 488. Notice the absence of specific labeling of the cell surface with antibodies against integrin α4, α5, and β5. Bar, 25 μm. Nuclei are stained using DAPI (blue). Representative images from 3 cornea explants are shown.

Article Snippet: The integrin-specific antibodies used for flow cytometry, in situ immunohistochemistry, colocalization, binding, and infection competition analyses were the following: MAbs anti-α2 (clone P1E6 [mouse]), anti-α3 (clones P1B5 [mouse] and ASC-1 [mouse]), anti-α4 (clones P4C2 [mouse] and PS/2 [rat]), and anti-α5 (clone P1D6 [mouse]), all from Merck Millipore; anti-α6 MAbs (clones GoH3 [rat] and MP 4F10 [mouse]) from Abcam; MAb anti-αV (clone 272-17E6 [mouse]) from Thermo Fisher Scientific; MAb anti-β1 (clone P5D2 [mouse]) and PAb anti-β1 (AF1778 [goat]), both from R&D Systems; MAb anti-β3 (clone MHF4 [mouse]) from Abnova; MAb anti-β4 (clone 422325 [mouse]) from R&D Systems; and PAb anti-β5 (H00003693D01P [rabbit]) from Abnova.

Techniques: Labeling, Staining

Journal: Journal of Virology

Article Title: Human Adenovirus Type 37 Uses α V β 1 and α 3 β 1 Integrins for Infection of Human Corneal Cells

doi: 10.1128/JVI.02019-16

Figure Lengend Snippet: Flow cytometry analysis of integrin subunit expression on HCE cells. The following antibodies were used to detect respective integrin subunits: α2 (P1E6), α3 (P1B5), α4 (P4C2), α5 (P1D6), α6 (GoH3), αV (272-17E6), β1 (P5D2), β3 (MF4), β4 (422325), and β5 (H00003693-D01P). Red and green colors represent staining in the presence and absence of primary antibody, respectively. n = 2.

Article Snippet: The integrin-specific antibodies used for flow cytometry, in situ immunohistochemistry, colocalization, binding, and infection competition analyses were the following: MAbs anti-α2 (clone P1E6 [mouse]), anti-α3 (clones P1B5 [mouse] and ASC-1 [mouse]), anti-α4 (clones P4C2 [mouse] and PS/2 [rat]), and anti-α5 (clone P1D6 [mouse]), all from Merck Millipore; anti-α6 MAbs (clones GoH3 [rat] and MP 4F10 [mouse]) from Abcam; MAb anti-αV (clone 272-17E6 [mouse]) from Thermo Fisher Scientific; MAb anti-β1 (clone P5D2 [mouse]) and PAb anti-β1 (AF1778 [goat]), both from R&D Systems; MAb anti-β3 (clone MHF4 [mouse]) from Abnova; MAb anti-β4 (clone 422325 [mouse]) from R&D Systems; and PAb anti-β5 (H00003693D01P [rabbit]) from Abnova.

Techniques: Flow Cytometry, Expressing, Staining

Journal: Journal of Virology

Article Title: Human Adenovirus Type 37 Uses α V β 1 and α 3 β 1 Integrins for Infection of Human Corneal Cells

doi: 10.1128/JVI.02019-16

Figure Lengend Snippet: Effects of preincubating HCE cells with anti-integrin antibodies on HAdV-37 and HAdV-5 infection and binding. (A) Effect of antibodies against integrin α subunits on HAdV-37 and HAdV-5 infection. (B) Effect of antibodies against integrin β subunits on HAdV-37 and HAdV-5 infection. (C) Effect of antibodies against integrin α and β subunits on HAdV-37 and HAdV-5 binding. Infection was assayed by counting virus-positive cells by immunofluorescence, and binding was assayed by quantitating 35 S-labeled virus association with cells. The results are presented as percentages of the control (i.e., untreated cells). The following antibodies were used: α2 (P1E6), α3 (P1B5), α4 (P4C2), α5 (P1D6), α6 (GoH3), αV (272-17E6), β1 (P5D2), β3 (MHF4), β4 (422325), and β5 (H00003693-D01P). The anti-GD1a specific MAb EM9 was used as a positive control. Results are shown as a percentage of the value for the control. *, P < 0.05; **, P < 0.01; ***, P < 0.001. n = 3.

Article Snippet: The integrin-specific antibodies used for flow cytometry, in situ immunohistochemistry, colocalization, binding, and infection competition analyses were the following: MAbs anti-α2 (clone P1E6 [mouse]), anti-α3 (clones P1B5 [mouse] and ASC-1 [mouse]), anti-α4 (clones P4C2 [mouse] and PS/2 [rat]), and anti-α5 (clone P1D6 [mouse]), all from Merck Millipore; anti-α6 MAbs (clones GoH3 [rat] and MP 4F10 [mouse]) from Abcam; MAb anti-αV (clone 272-17E6 [mouse]) from Thermo Fisher Scientific; MAb anti-β1 (clone P5D2 [mouse]) and PAb anti-β1 (AF1778 [goat]), both from R&D Systems; MAb anti-β3 (clone MHF4 [mouse]) from Abnova; MAb anti-β4 (clone 422325 [mouse]) from R&D Systems; and PAb anti-β5 (H00003693D01P [rabbit]) from Abnova.

Techniques: Infection, Binding Assay, Virus, Immunofluorescence, Labeling, Control, Positive Control

Journal: Journal of Virology

Article Title: Human Adenovirus Type 37 Uses α V β 1 and α 3 β 1 Integrins for Infection of Human Corneal Cells

doi: 10.1128/JVI.02019-16

Figure Lengend Snippet: Effect of penton base-derived, RGD-containing peptides (4 mM) on HAdV-5 and HAdV-37 infection of (A) and binding to (B) HCE cells. 37-RGD, peptide mimicking RGD-containing loop of HAdV-37 penton base. 37-AAA, RGD-containing loop in HAdV-37 penton base peptide replaced with AAA. GRGDSP, peptide used previously to inhibit HAdV infection of human cells. The results are presented as a percentage of the value for the control (i.e., untreated cells). *, P < 0.05; **, P < 0.01; ***, P < 0.001. n = 3.

Article Snippet: The integrin-specific antibodies used for flow cytometry, in situ immunohistochemistry, colocalization, binding, and infection competition analyses were the following: MAbs anti-α2 (clone P1E6 [mouse]), anti-α3 (clones P1B5 [mouse] and ASC-1 [mouse]), anti-α4 (clones P4C2 [mouse] and PS/2 [rat]), and anti-α5 (clone P1D6 [mouse]), all from Merck Millipore; anti-α6 MAbs (clones GoH3 [rat] and MP 4F10 [mouse]) from Abcam; MAb anti-αV (clone 272-17E6 [mouse]) from Thermo Fisher Scientific; MAb anti-β1 (clone P5D2 [mouse]) and PAb anti-β1 (AF1778 [goat]), both from R&D Systems; MAb anti-β3 (clone MHF4 [mouse]) from Abnova; MAb anti-β4 (clone 422325 [mouse]) from R&D Systems; and PAb anti-β5 (H00003693D01P [rabbit]) from Abnova.

Techniques: Derivative Assay, Infection, Binding Assay, Control

Journal: Journal of Virology

Article Title: Human Adenovirus Type 37 Uses α V β 1 and α 3 β 1 Integrins for Infection of Human Corneal Cells

doi: 10.1128/JVI.02019-16

Figure Lengend Snippet: HAdV-37 virions colocalize with α3 and αV integrins on HCE cells. Alexa Fluor 555-labeled HAdV-37 virions (in orange) were incubated with HCE cells at 37°C for different time points and subsequently stained for α3 (P1B5) or αV (272-17E6) integrins (in green). (A) Representative confocal images of HAdV-37 virions colocalizing with integrin antibodies after 15 min (top, α3; bottom, αV). Slices 90° tilted show optical sections of the x-z and y-z planes at the bottom and right side, respectively, of each image on the left. The marked squares display a colocalization event, enlarged to the right in the center of the crosshairs in each box and in the x-y , x-z , and y-z sections. Scale bars, 5 μm. (B) Quantification of the percentage of colocalized HAdV-37 virions with α3 and αV integrins at different time points in HCE cells. Data were collected from 5 to 7 z stacks with at least 2 cells per stack. #, Bars represent data from a pixel shift analysis to test if colocalization occurs by chance. *, P < 0.05. n = 1.

Article Snippet: The integrin-specific antibodies used for flow cytometry, in situ immunohistochemistry, colocalization, binding, and infection competition analyses were the following: MAbs anti-α2 (clone P1E6 [mouse]), anti-α3 (clones P1B5 [mouse] and ASC-1 [mouse]), anti-α4 (clones P4C2 [mouse] and PS/2 [rat]), and anti-α5 (clone P1D6 [mouse]), all from Merck Millipore; anti-α6 MAbs (clones GoH3 [rat] and MP 4F10 [mouse]) from Abcam; MAb anti-αV (clone 272-17E6 [mouse]) from Thermo Fisher Scientific; MAb anti-β1 (clone P5D2 [mouse]) and PAb anti-β1 (AF1778 [goat]), both from R&D Systems; MAb anti-β3 (clone MHF4 [mouse]) from Abnova; MAb anti-β4 (clone 422325 [mouse]) from R&D Systems; and PAb anti-β5 (H00003693D01P [rabbit]) from Abnova.

Techniques: Labeling, Incubation, Staining

Journal: Journal of Virology

Article Title: Human Adenovirus Type 37 Uses α V β 1 and α 3 β 1 Integrins for Infection of Human Corneal Cells

doi: 10.1128/JVI.02019-16

Figure Lengend Snippet: HAdV infection (in green) and expression of integrin subunits α3, αV, and β1 (in red) on HCE cells grown as multilayers at the air-liquid interphase. HAdV-5 and -37 were infected from the apical (upper) side. Antibodies used for staining were P1B5 (α3), 272-17E6 (αV), P5D2 (β1), and B025/AD51 (adenovirus hexon). Nuclear stain is shown in blue. Staining with secondary antibodies only was completely negative. White bar, 50 μm. n = 2.

Article Snippet: The integrin-specific antibodies used for flow cytometry, in situ immunohistochemistry, colocalization, binding, and infection competition analyses were the following: MAbs anti-α2 (clone P1E6 [mouse]), anti-α3 (clones P1B5 [mouse] and ASC-1 [mouse]), anti-α4 (clones P4C2 [mouse] and PS/2 [rat]), and anti-α5 (clone P1D6 [mouse]), all from Merck Millipore; anti-α6 MAbs (clones GoH3 [rat] and MP 4F10 [mouse]) from Abcam; MAb anti-αV (clone 272-17E6 [mouse]) from Thermo Fisher Scientific; MAb anti-β1 (clone P5D2 [mouse]) and PAb anti-β1 (AF1778 [goat]), both from R&D Systems; MAb anti-β3 (clone MHF4 [mouse]) from Abnova; MAb anti-β4 (clone 422325 [mouse]) from R&D Systems; and PAb anti-β5 (H00003693D01P [rabbit]) from Abnova.

Techniques: Infection, Expressing, Staining