Figure 2 - Jund expression and protein levels in the NTN-susceptible WKY rat.

(a,b) Microarray (a) and QRT-PCR (b) analyses showed that Jund expression is higher in WKY than in LEW glomeruli at baseline (WKY_c, LEW_c) and in NTN-induced glomeruli (WKY_NTN, and LEW_NTN). At least three rats per strain and per condition (control or NTN) were used. Error bars, s.e.m. (c) Immunostaining for JunD protein in WKY and Lewis glomeruli after NTN induction (day 10), showing increased JunD in the WKY glomeruli. (d) Sequence analysis of the rat Jund promoter, showing a C/T polymorphism at - 210 bp (asterisk) in the vicinity of an octamer binding motif (- 219 to - 212 bp). (e) Luciferase assay performed after transfecting COS7 cells with pGL3-Basic vector containing, 300 bp upstream of the transcription initiation site, either the WKY or LEW Jund promoters (pGL3-WKY and pGL3-LEW, respectively). Firefly luciferase activity, normalized to Renilla luciferase activity, is expressed relative to the activity of the empty pGL3-Basic vector. Error bars, s.e.m. for five different transfection experiments performed in replicates of six. (f) JunD binding to AP-1 site in BMDMs. Specific JunD binding to AP-1 consensus sequence nucleotides (5'-TGAGTCA-3') was greater in WKY BMDM nuclear extracts than in LEW and WKY.LCrgn2 by TransAM assay. A₄₅₀, absorbance at 450 nm. ^*P < 0.05; error bars, s.e.m.