cd151 specific mouse mab 11g5a  (Bio-Rad)

Journal: Antimicrobial Agents and Chemotherapy

Article Title: HSPG-binding peptide Pep19-2.5 is a potent inhibitor of HPV16 infection

doi: 10.1128/aac.01575-24

Figure Lengend Snippet: Analysis of CD151 crowding at PsV-binding sites. ( A ) HeLa cells were incubated for a total of 4 hours with PsVs. After 1 hour, they were additionally incubated either without any peptide (−pep.), with 1 µg/mL of the control peptide (ctrl.), or with Pep19-2.5 (pep.). Then, cells were fixed and surface-stained for L1 (magenta) and CD151 (green). x , y -scans of L1 in the confocal and CD151 in the STED channel were recorded at the basal membrane. Orange boxes mark enlarged views of the overlays shown to the right. ( B ) Analysis of the normalized L1 neighbored CD151 maxima (for details see Materials and Methods). At every L1 position, a circular 925 nm ROI was positioned, the CD151 maxima were counted, and the value was normalized to the CD151 average maxima density. The number of PsVs (defined by the L1 signal) is plotted in absolute counts vs the normalized CD151 maxima counts. At a randomized relationship between L1 and CD151 (horizontally and vertically flipped images), the analysis is supposed to yield a Gaussian-like distribution is supposed to peak at the value of 1. If CD151 maxima at PsV locations are more abundant than random, the distribution becomes right shifted, as it contains two Gaussian distributions. Upper panel: original data showing right-shifted distributions. For determining the fraction of PsVs locating at CD151 crowds, the distribution was decomposed into two Gaussian distributions (dotted lines mark randomized, dashed lines mark the pool of PsVs at accumulated CD151 (crowds), and gray dotted-dashed lines are the sum of both populations). The size of the crowded fractions ranges on average between 21% and 34%. Lower panel: flipped data. The fitted Gaussians peak at 1. The size of the crowded fractions ranges on average between 1% and 7%. ( C ) Same data as in ( B ), showing the average of the normalized L1 neighbored CD151 maxima. Statistical differences between the groups ( n = 3) were analyzed using Welch’s t test ( P = 0.0027 for −pep. original vs flipped, P = 0.0232 for ctrl. original vs flipped, P = 0.0051 for pep. original vs flipped, P = 0.8453 for ctrl. vs pep. original, P = 0.7432 for ctrl. vs pep. original, P = 0.9096 for ctrl. vs pep. flipped, P = 0.5371 for ctrl. vs pep. flipped). ( D ) Pearson correlation coefficient (PCC) in the ROI between the L1 and CD151 original and flipped (randomized) images. Statistical differences between the groups ( n = 3) were analyzed using Welch’s t test ( P = 0.0042 for −pep. original vs flipped, P = 0.0034 for ctrl. original vs flipped, P = 0.0015 for pep. original vs flipped, P = 0.6470 for ctrl. vs pep. original, P = 0.2136 for ctrl. vs pep. original, P = 0.9628 for ctrl. vs pep. flipped, P = 0.6924 for ctrl. vs pep. flipped). (B, C) Values are given as mean ± standard deviation (SD) ( n = 3 from three biological replicates, including 15 images per condition and replicate).

Article Snippet: CD151-specific mouse mAb (11G5a) was obtained from Bio-Rad (Munich, Germany). β-Actin-specific mouse mAb (A5441) was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Binding Assay, Incubation, Control, Staining, Membrane, Standard Deviation

Journal: Antimicrobial Agents and Chemotherapy

Article Title: HSPG-binding peptide Pep19-2.5 is a potent inhibitor of HPV16 infection

doi: 10.1128/aac.01575-24

Figure Lengend Snippet: The Pep19-2.5 induces large extracellular PsV accumulations. ( A ) Same preparation as in . In each condition, we imaged by confocal microscopy employing x , z -scans /y , z -scans those cells exhibiting the most prominent PsV accumulations. Shown are images of the CD151 and L1 channels of the different conditions (same scaling of the respective channels). We show two examples of the Pep19-2.5 condition to point out that, in the presence of Pep19-2.5, we observe cells resembeling those in the control condition, as well as cells with very large PsV accumulations, which are typically localized at the top of the cell (see also ). ( B ) Same images as in ( A ), auto-scaled, and shown as overlays (L1 in magenta and CD151 in green) with magnified views of the orange boxes. Values in the upper right indicate the fraction of distal PsVs from which in ( C ) we show the average values. ( C ) Cells were outlined (for example, see ) and the background-corrected L1 signal within the outline was subtracted from the total background-corrected L1 signal, yielding the signal of distal PsVs. For details regarding the calculation of the fraction of distal PsVs, see Materials and Methods. Values are given as means ± SD ( n = 3 from three biological replicates, including 20 images per condition and replicate; for one replicate, we show almost the complete data set in ). Statistical differences between the groups ( n = 3) were analyzed using Welch’s t test ( P = 0.3033 for ctrl. vs pep., P = 0.0348 for ctrl. vs pep., P = 0.0167 for −pep. vs pep.).

Article Snippet: CD151-specific mouse mAb (11G5a) was obtained from Bio-Rad (Munich, Germany). β-Actin-specific mouse mAb (A5441) was obtained from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Confocal Microscopy, Control