Generation of BALB/cByJ-CD11B-Kif1cD2 transgenic (Tg-Kif1cD2) mice.
(A) Schematic representation of the Kif1c gene used to generate the transgenic mice showing the promoter (CD11B/ITGAM), and the Kif1c gene, followed by the hGH/polyA signal sequence. Arrows indicate PCR-primers for screening. (B and C) Percentage of splenic F4/80+ (B) and CD11b+ (C) cells of Tg-Kif1cD2 and NLC. The analysis was performed on gated live cells according to their FSC vs. SSC profile. Statistical significance was determined using the Mann-Whitney U test. Data represent the mean ± SEM of at least 5 individual mice. (D) Kif1c expression in thioglycolate-induced adherent cells by Western blotting using whole-cell extracts and the anti-Kif1c mAb. Actin was used as a loading marker. (E) mRNA expression of Kif1c was measured from sorted TCRβ−CD19−CD11b+ myeloid cells of CByJ mice and compared with TCRβ−CD19−CD11b+ myeloid cells of D2 mice. β2-microglobulin and GAPDH were used as an endogenous control. Data represent the mean ± SEM of two experiments (pool of 5 animals/each).