OSU-T315 as an Interesting Lead Molecule for Novel B Cell-Specific Therapeutics

Figure 1

Identification of mice with B cell-specific deletion of ILK. Mice with B cell-specific deletion of ILK were identified through PCR (a) and flow cytometry (b) and Western blot (c). (a) DNA from the tail was amplified by PCR and then separated and visualized on 2% agarose gel to identify the mice with floxed ILK genes: L: TrackIt™ 100 bp DNA ladder; WT: ILK^WT/WT; H: ILK^flox/WT; and F: ILK^flox/flox. (b) Identification of mice with CD19^WT/WT, CD19^Cre/WT, or CD19^Cre/Cre genotype was done by flow cytometric analysis on the blood that was double-stained with CD19 (FITC) and CD45R/B220 (PE/Cy5). Mice with CD19^Cre/Cre genotype did not display CD19 on the outer surface of the B cells. CD19^Cre/WT mice still expressed CD19 on the outer surface of the B cells, but at a lower density than CD19^WT/WT mice which consequently translated in a lower MFI compared to CD19^WT/WT mice. The similar percentages of gated CD45R/B220⁺ cells with equal MFI demonstrated that there was no loss in the B cell population in CD19^Cre/WT mice and CD19^Cre/Cre mice compared to CD19^WT/WT mice. (c) Western blot was performed on splenocytes and on splenic B cells from mice identified as being ILK wild type (WT, CD19^WT/WT/ILK^flox/flox) or ILK knockout (KO, CD19^Cre/WT/ILK^flox/flox). High expression of ILK was observed in the splenocytes of WT and KO mice, but ILK was not detected in the B lymphocytes of the latter.

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