Fractalkine: An important candidate for directing periglomerular leukocyte accumulation in irradiated mouse kidneys

Figures & data

Table I.  Specific primary antibodies against proteins of interest.

Protein of interest	Primary antibody	Catalog #
Fractalkine (CX3CL1)	Goat-anti rat anti Fractalkine (R&D)	AF537
Leukocytes (CD45)	Biotinylated rat anti-mouse anti-CD45	Old#: 01112D
	(PharMingen)	New#: 553078
PECAM-1 (CD31)	Purified rat anti-mouse anti CD31	Old#: 01951D
	(PharMingen)	New#: 553370

Figure 1.  Immunohistochemical staining patterns of fractalkine. (A) ring formation, (B) surface coverage <30% and (C) surface coverage >30%.

Table II.  Primersequences.

Gene of interest	Primersequence
Fractalkine	Forward 5′-GGGTGGCCATGTTTGCTTAC-3′
	Reverse 5′-CAGGCAAGCAGCTCACACTG-3′
PECAM-1	Forward 5′-GAGCCCAATCACGTTTCAGTTT-3′
	Reverse 5′-TCCTTCCTGCTTCTTGCTAGCT-3′

Figure 2.  Photographs of mouse kidney sections stained immunohistochemically for leukocytes (A–B), fractalkine (C–D) and PECAM-1 (E–F) at 40 weeks after irradiation with 0 Gy or 16 Gy. All photomicrographs were taken at an original magnification of 400×. Sections were counterstained with hematoxylin and eosin.

Figure 3.  Semi-quantitative analysis of the number of fractalkine positive glomeruli as a percentage of the total number of glomeruli at different time intervals after irradiation or sham-treatment. Glomeruli were evaluated separately with surface coverage ranging from ring formation to < or >30%. Data are expressed as mean±S.D.

Figure 4.  Relative mRNA expressions of genes encoding fractalkine (A) and PECAM-1 (B). Messenger RNAs were analyzed by real-time PCR assays and normalized for GAPDH, a housekeeping gene.