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Table 1 Antibody panels used for FCM analyzes

From: PRRSV-infected monocyte-derived dendritic cells express high levels of SLA-DR and CD80/86 but do not stimulate PRRSV-naïve regulatory T cells to proliferate

Antigen

Clone

Isotype

Fluorochrome

Labeling strategy

Source of primary Ab

Phenotyping of moDCs

CD1

76-7-4

IgG2a

Alexa647

secondary antibodya

in housej

CD14

CAM36A

IgG1

BV421

secondary antibodyb

VMRD

CD80/86

CD152/Fc chimera

IgG2a

Alexa647

secondary antibodya

Sigma-Aldrich

CD163

2A10/11

IgG1

PE

directly conjugated

AbD Serotec

CD169

3B11/11

IgG1

BV421

secondary antibodyb

AbD Serotec

CD172a

74-22-15A

IgG2b

Alexa488

secondary antibodyc

in housej

SLA-DR

MSA3

IgG2a

Alexa647

secondary antibodya

in housek

Assessment of moDC infection rate

CD172a

74-22-15A

IgG2b

PE

secondary antibodyd

in housej

PRRSV-N

P10/b1

IgG1

Alexa488

directly conjugatede

in housel

CD163, CD169, SLA-DR and CD80/86 expression on infected moDCs

CD172a

74-22-15A

IgG2b

Alexa647

secondary antibodyf

in housej

SLA-DR

MSA3

IgG2a

PE

directly conjugatedg

in housek

CD80/86

hCTLA4/Fc chimera

IgG2a

BV421

two step biotin-streptavidinh

Sigma-Aldrich

CD163

2A10/11

IgG1

PE

directly conjugated

AbD Serotec

CD169

3B11/11

IgG1

BV421

two step biotin-streptavidini

AbD Serotec

PRRSV-N

P10/b1

IgG1

Alexa488

directly conjugatede

in housel

  1. aGoat anti-Mouse IgG2a-Alexa647, Life Technologies.
  2. bRat anti-Mouse IgG1-BV421, clone RMG1-1, BioLegend.
  3. cGoat anti-Mouse IgG2b-Alexa488, Life Technologies.
  4. dGoat anti-Mouse IgG2b-PE, Southern Biotech.
  5. eAlexa Fluor® 488 Protein Labeling Kit, Life Technologies.
  6. fGoat anti-Mouse IgG2b-Alexa647, Life Technologies.
  7. gLightning-Link™ RPE Conjugation Kit, Innova Biosciences.
  8. hGoat anti-Mouse IgG2a-biot., Southern Biotech; Streptavidin-BV421, BioLegend.
  9. iGoat anti-Mouse IgG1-biot., Southern Biotech; Streptavidin-BV421, BioLegend.
  10. jPescovitz et al. [27].
  11. kHammerberg and Schurig [28].
  12. lWeiland et al. [29].