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. 2021 Oct 28;220(12):e202005184. doi: 10.1083/jcb.202005184

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© 2021 Kumari et al.

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Figure 4. — Pin1 interacts with the Hook2-Nde1-Lis1 dynein complex, but not with the spindly-dynactin-dynein complex. (A) Immunoblots showing affinity precipitates of GST-Pin1/GST from prometaphase (nocodazole-treated) and metaphase (released into MG132 after nocodazole treatment) HeLa cell lysates, probed for the indicated antigens. (B) Immunoblots showing the affinity precipitates of GST-Pin1 from SST-MTAP and AAA-MTAP prometaphase U2OS stable cell lysates, probed for the indicated antigens. (C and D) Ratios (AAA-MTAP/SST-MTAP) of eluate (beads) fraction band intensities for each indicated antigen (x axes) over three independent affinity purification (AP) experiments. (E) Representative single confocal z-sections of SST- or AAA-MTAP U2OS stable cells showing the localization of the indicated antigens. Boxes are magnified in the insets. The metaphase images are deconvolved to enhance spindle fiber visualization. Arrows indicate colocalization. Scale bar = 10 µm, inset scale bar = 5 µm. IB, immunoblot. Error bars = mean ± SEM. ***, P < 0.001; ****, P < 0.0001 (one-way ANOVA).