Figure 1. The expression of E3 ubiquitin... | Rockefeller University Press

Figure 1.

$Figure 1. The expression of E3 ubiquitin ligases and the deletion of Mib1 in the lymphoid tissues. (A and B) The expression of E3 ligases in the thymus (A) and spleens (B). Total RNAs were prepared from the thymus and spleen (total) and nonhematopoietic (CD45−) and hematopoietic (CD45+) fractions sorted from collagenase-treated thymi and spleens. They were subjected to quantitative real-time RT-PCR using specific primers for Cd45, Mib1, Mib2, Neur1, and Neur2. Data are mean ± SD from triplicate experiments. The mean expression level of Mib1 mRNA in the total thymus and spleen was defined as 1 arbitrary unit, as compared with the expression levels of the E3 ubiquitin ligases. (C) Immunohistochemistry was performed on the cryosections from the MMTV-Cre;Mib1+/f and MMTV-Cre;Mib1f/f thymi with the anti-Mib1 antibody. Note that Mib1 expression (arrowheads) was reduced in the MMTV-Cre;Mib1f/f mice thymus. Bars, 5 μm. (D) Total protein extracts of the spleens from the MMTV-Cre;Mib1+/f and MMTV-Cre;Mib1f/f mice were immunoblotted with the anti-Mib1 antibody.$

The expression of E3 ubiquitin ligases and the deletion of Mib1 in the lymphoid tissues. (A and B) The expression of E3 ligases in the thymus (A) and spleens (B). Total RNAs were prepared from the thymus and spleen (total) and nonhematopoietic (CD45⁻) and hematopoietic (CD45⁺) fractions sorted from collagenase-treated thymi and spleens. They were subjected to quantitative real-time RT-PCR using specific primers for Cd45, Mib1, Mib2, Neur1, and Neur2. Data are mean ± SD from triplicate experiments. The mean expression level of Mib1 mRNA in the total thymus and spleen was defined as 1 arbitrary unit, as compared with the expression levels of the E3 ubiquitin ligases. (C) Immunohistochemistry was performed on the cryosections from the MMTV-Cre;Mib1⁺^/f and MMTV-Cre;Mib1^f/f thymi with the anti-Mib1 antibody. Note that Mib1 expression (arrowheads) was reduced in the MMTV-Cre;Mib1^f/f mice thymus. Bars, 5 μm. (D) Total protein extracts of the spleens from the MMTV-Cre;Mib1⁺^/f and MMTV-Cre;Mib1^f/f mice were immunoblotted with the anti-Mib1 antibody.