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. 2010 Nov 8;5(11):e15383. doi: 10.1371/journal.pone.0015383

Figure 1. Expression of NIKΔT3 induced by both catK-Cre and lysM-Cre.

Figure 1

A. Diagram of NT3 locus. NIKΔT3 (NT3) cDNA was knocked into the ROSA26 locus, flanked upstream by a loxP- NeoR.STOP-loxP cassette. Cre-mediated loxP recombination removes the NeoR.STOP sequence, putting NT3 under control of the ROSA 26 promoter. Black arrows indicate the pair of primers used to amplify across the loxP. B–C. Genomic DNA amplification across the loxP sites using NT3.catK (B) and NT3.lysM (C) BMMs cultured with 20 ng/ml RANKL for up to 4 days. ∼3kb STOP and ∼300bp lox bands are generated before and after Cre-mediated deletion, respectively. The left most lanes (-) are negative controls with no genomic DNA in PCR reactions. D–E. Western blot analysis of NT3 expression in total cell lysates, using BMMs cultured as in B. NIK KO BMMs serve as a negative control. F–G. p100/p52 processing was assessed by Western blot in total cell lysates from Ctl and NT3.catK BMMs cultured in RANKL for the indicated times, or in unstimulated NT3.lysM BMMs. H. Evaluation of RelB and p65 levels in nuclear extracts of NT3.lysM BMMs. I. Western blot analysis of IκBα phosphorylation in NT3.lysM BMM total lysates.