CUL3 ^KLHL12 is needed for the accumulation of Akita condensates at junctions. (A) Quantitation of LNPK-GFP puncta in LAMP1-mCherry structures in siCtrl and siRNA-treated cells incubated with Baf (6 h). Representative images on the left and in Fig. S9 F. The control for each condition was set to 1.0. (B) Quantitation of Akita-sfGFP puncta in LAMP1-mCherry structures in siCtrl and siRNA-treated cells with Baf (3.5 h). Representative images on the left and in Fig. S9 G(C) Images and the percent cells with large Akita puncta (≥0.5 µm²) (left), and the mean size of Akita puncta (right) in Ctrl and siRNAi-treated cells. Arrows mark large Akita puncta. (D) Left, images of cells transfected with Akita-sfGFP and LNPK-mCherry in siCtrl or siCUL3-treated cells. Arrowheads mark Akita puncta that colocalize with LNPK puncta. Quantitation is on the right. Error bars represent SEM, n = 3 independent experiments. The results were quantified from 62 to 78 cells in A, 59–77 cells in B, 112–154 cells in C, and 39–44 cells in D. NS: not significant (P ≥ 0.05), * (P < 0.05), **(P < 0.01), Student’s unpaired t test.