Figure 5: PE requires SUN1/2 and the translocon complex for transport to the nucleoplasm.
From: Nuclear envelope-associated endosomes deliver surface proteins to the nucleus
(a) Flowchart and (b) results of siRNA screening; PE and Nu-T detection as in Fig. 4. Representative of five different replicates. (c–e) MG63 cell fractionation after Sec61B or SUN2 knockdown. (c) Fractionation controls (as in Fig. 4). (d) PE subcellular distribution in MG63 cells after Sec61B and SUN2 knockdown. Representative of three independent repeats. (e) Quantification of PE in nuclear fraction after Sec61B and SUN2 knockdown. Error bars at s.d. t-Test type 2, two tails. (f) Co-labelling of PE with EGFR. Nucleus is delimited by yellow circle. Scale bar, 10 μm. (g) A431 cells fractionation. (h) Input loading control of A431 protein extract. (i) EGFR nuclear distribution in A431 cells after Sec61B and SUN2 knockdown. Representative of three independent repeats. (j) Quantification of EGFR in nuclear fraction. Error bars at s.d. t-Test type 2, two tails. (k) Two intracellular routes of cargos. After endocytosis, NAE dock on the nucleus, discharging their content into the nucleoplasm in a SUN1/2- and Sec61 translocon-dependent manner.
