Fig. 1: Generation of fully human Ab-producing mice.
a Representation of the IGHK-NAC structure. Human Ig heavy chain locus (IGH) derived from hChr.14 and Ig kappa light chain locus (IGK) derived from hChr.2 are represented on a mouse artificial chromosome (MAC). b Representative image of metaphase FISH analysis with IGK-BAC (CH17-216K2) (red) and IGH-BAC (CH17-212P11) (green) detecting the IGHK-NAC in bone marrow cells from a TC-mAb mouse. Arrow indicates the IGHK-NAC and the inset shows an enlarged image thereof. Scale bar (10 μm). c GFP images of different tissues from a Tc mouse carrying the IGHK-NAC. GFP expression indicates the presence of the IGHK-NAC. BF, bright field. Scale bar (5 mm). d Retention rate of the IGHK-NAC in various Tc mouse tissues analysed by FISH. Li liver, In intestine, Ki kidney, Sp spleen, Lu lung, He heart, Sk skeletal muscle, Thy thymus, Br brain, Ut uterus. e Percentage of GFP-expressing cells in peripheral blood mononuclear cells from Tc mice of different generations. f Percentage of B220-positive cells in the lymphocyte fraction of peripheral blood in age-matched WT (n = 10), hetero HKLD (n = 10, Igh+/−, Igk+/− and Igl1+/low), HKLD (n = 10, Igh−/−, Igk−/− and Igl1low/low) and TC-mAb mice (n=1106). Box plots are indicated in terms of minima, maxima, centre, bounds of box and whiskers (1.5 interquartile range value), and percentile in the style of Tukey (e, f). *P < 0.001 (unpaired Dunn’s test for multiple comparison). Gating strategies are presented in Supplementary Fig. 7. g RT-PCR analysis of total RNA from various tissues of the Tc mouse. Li liver, In intestine, Ki kidney, Sp spleen, Lu lung, He heart, Sk skeletal muscle, Thy thymus, Br brain, Ut uterus. M size marker. Twelve-week-old (b) and 14-week-old female TC-mAb mice (c, d, g) were analysed. Assessment of the GFP- and B220-positive ratios in lymphocytes by FCM was applied to 4–5-week-old TC-mAb mice (e, f). Five female and five male mice aged 4–5 weeks each were used for WT, hetero HKLD and HKLD (f).
