Skip to main content
. 2017 Jul 24;27(14):2123–2136.e7. doi: 10.1016/j.cub.2017.06.021

Figure 5.

Figure 5

PCM1 Specifically Regulates GABARAP Protein Levels through Its LIR Motif

(A) HEK293A cells treated with RF or PCM1 siRNA incubated in full medium (FM) or EBSS with or without BAFA for 2 hr.

(B) Quantifications from (A). For LC3-II/actin, n = 3. For p62/actin, n = 5. For NBR1/actin, n = 5. For PCM1/actin, n = 5. Mean ± SEM. One-way ANOVA; ∗p ≤ 0.05.

(C and D) HEK293A cells treated with RF or PCM1 siRNA incubated in FM or EBSS with or without BAFA1 for 2 hr. In (C), GABARAP-I and GABARAP-II are resolved, whereas in (D) only GABARAP-I is resolved in the western blot.

(E) Quantifications from (C) and (D). For GABARAP-II/actin, n = 5. For GABARAP-I/actin, n = 3. Mean ± SEM. One-way ANOVA; ∗p ≤ 0.05.

(F) HEK293A cells expressing the indicated constructs were subjected to immunoblot.

(G) Quantification of (F). For p62/actin, n = 4. For GABARAP-I/actin, n = 5. Mean ± SEM; unpaired Student’s t test; ∗∗p ≤ 0.01.

(H) HEK293A cells treated with RF or PCM1 siRNA (72 hr total) and transfected with the indicated siPCM1-resistant constructs (last 24 hr) and immunoblotted.

(I) Quantifications from (H). For RF+GFP, siPCM1+GFP, and siPCM1+GFP-PCM1 WT, six experiments are shown. For siPCM1+GFP-PCM1 3xAla, three experiments are shown. Mean ± SEM; unpaired Student’s t test; ∗∗p ≤ 0.01.