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. 2017 Jul 17;8:806. doi: 10.3389/fimmu.2017.00806

Figure 1.

Figure 1

Detection of Lip10 accumulation in RAJI B cell line upon cathepsin S-specific inhibition. (A) Schematic diagram of invariant chain (Ii) proteolytic removal as part of the major histocompatibility complex (MHC) class II presentation pathway. Inhibition of cathepsin S activity leads to an accumulation of cathepsin S substrate Lip10 in professional antigen-presenting cells. (B) Lip10 accumulation after cathepsin S inhibition was detected by western blot using an anti-CD74 antibody in RAJI cells incubated for 20 h with cathepsin S-specific inhibitor RO5459072. (C) Twelve antibody clones raised in rabbits against the C-terminal end of Lip10 were tested for the ability to detect intracellular Lip10 accumulation by staining RAJI cells incubated for 20 h with RO5459072 in a flow cytometry-based assay. Lip10 median fluorescence intensity (MFI) is displayed for DMSO-treated (red line) and RO5459072-treated (solid blue) RAJI cells. The stain index, calculated for each clone, with (MFI of cathepsin S inhibitor-treated cells − MFI of DMSO-treated cells)/SD of DMSO-treated cells, is displayed in parenthesis after each clone name.