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Blood, Vol. 110, Issue 12, 4077-4085, December 1, 2007
Kupffer cell heterogeneity: functional properties of bone marrow–derived and sessile hepatic macrophages
Blood Klein et al.
110: 4077
Supplemental materials for: Klein et al
Files in this Data Supplement:
- Figure S1. Controls of the immunohistological staining protocol (JPG, 61.1 KB)
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(A) Liver sections from naive B6.CD45.2 animals were stained with unconjugated F4/80 mAb which were consecutively labeled with a secondary Alexa 546-conjugated anti-rat antibody (red). Positively stained cells are abundant in the intra-sinusoidal compartment and display an elongated morphology characteristic of Kupffer cells. (B) Sections stained only with the secondary Alexa 546-conjugated anti-rat antibody without the primary F4/80 mAb showed no non-specific binding of the conjugated antibody. To exclude non-specific binding of the congenic mAbs anti-CD45.2 and anti CD45.1 in experimental bone marrow chimeras or liver transplant recipients, the immunohistological staining protocol (F4/80 + secondary Alexa 546 mAb; anti-CD45.2 FITC and anti-CD45.1 Cy5 – displayed in false color blue) was used in naive B6.CD45.2 (C) and B6.CD45.1 (D). CD45.1-positive Kupffer cells appear in purple, whereas CD45.2-positive Kupffer cells are orange. There was no non-specific CD45.1 staining in B6.CD45.2 mice (C) and no non-specific CD45.2 staining in B6.CD45.1 animals (D). This set of positive and negative staining controls validates the immunohistologic technique that was used to identify persistent “sessile” Kupffer cells in bone-marrow chimeras (Figure 2) and liver transplant recipients (Figure 7).
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