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. 2023 May 11;299(6):104811. doi: 10.1016/j.jbc.2023.104811

Figure 2.

Figure 2

The kinetics of proteasome catalytic β subunits are measurable using MV151 across tissue homogenates.A, top, lysates from mouse cortical tissue were incubated with MV151, and samples were collected at indicated time points. Samples were applied to SDS-PAGE and imaged with the Cy3 channel to detect MV151 labeling. MV151 bands correspond to individual catalytically active β subunits (arrows). Bottom, MV151 gels were immunoblotted with antibodies to the indicated core proteasome subunits (β2, β5) and actin. β2 and β5 immunoblots show unmodified subunit (arrow) and ∼1.1 kDa MV151-shifted subunit (∗∗). B, quantification of actin-normalized MV151 signals from (A). Data points are represented as mean ± SEM (n = 3). C, top, Cy3 in-gel visualization of MV151 time course with lysates from mouse liver tissue treated as in (A). Bottom, representative immunoblots of the MV151 gel as in (A). D, quantification of actin-normalized MV151 signals from (C). Data points are represented as mean ± SEM (n = 3). E, comparison of quantification of MV151 signal from mouse cortex (B) and mouse liver (D). Data are represented as mean ± SEM (n = 3). Statistical test: two-way ANOVA (∗p < 0.0332, ∗∗p < 0.0021). See also Fig. S2.