Figure S5. EPG-7 mediates autophagic degradation of several other types of protein aggregates and is dispensable for starvation-induced autophagic degradation of substrate proteins. (A-D) Loss of function of
c17e4.2 (A and B) or
w07g4.5 (C and D) does not cause a defect in degradation of SQST-1. No SQST-1 aggregates are detected in any stages of embryos. (A and C) DAPI images of the embryos shown in B and D, respectively. (E) C33D9.6::GFP is weakly expressed and diffusely localized in the cytoplasm of early stage embryos. (F) At larval stages, C33D9.6::GFP is largely diffusely localized apart from a few aggregates in hypodermal cells. (G) At the post-embryonic stage, C17E4.2::GFP is localized in the intestine and forms a few aggregates. (H) At larval stages, W07G4.5::GFP forms aggregates that are localized in the cytoplasm and also in the nuclei (blue) in intestinal cells. (I and J) In wild-type embryos, C33D9.6::GFP is weakly expressed and diffusely localized in the cytoplasm apart from a few punctate structures. (I) DIC image of the embryo shown in J. (K and L) C33D9.6:: GFP is present at dramatically increased levels and forms a large number of aggregates in
atg-3 (K) and
epg-7 (L) mutant embryos. (M) Separate images for Fig. 7 J. C17E4.2::GFP aggregates colocalize with EPG-7 aggregates in
atg-3 mutants. Insets show a magnified view. (N and O) C33D9.6::GFP punctate structures overlap with EPG-7 aggregates (N) but are separable from SQST-1 aggregates (O) in atg- 3 mutants. (P and Q) In
atg-3 mutants, W07G4.5 punctate structures are largely colocalize with EPG-7 aggregates (P) but separable from SQST-1 aggregates (Q). (R and S) In wild-type embryos, a large number of SEPA-1 aggregates are present at the 100-cell stage. (R) DAPI image of the embryo shown in S. (T and U) A large number of SEPA-1 aggregates accumulate in
let-363 mutant embryos, derived from
let-363(
ok3018) I/hT2[
bli-4(
e937) let- ?(
q782) qIs48](I;III). (T) DAPI image of the embryo shown in U. (V-X) SQST-1 is weakly expressed in wild-type embryos (V and W) and its expression level remains unchanged in
rheb-1(RNAi) embryos (X). (Y) SQST-1 accumulates into a large number of aggregates in atg- 18 mutant embryos. (Z) SQST-1 forms aggregates in
epg-7 mutant embryos. (A2) Expression of SQST-1 in
epg-7 mutant embryos is not affected by inactivation of
let-363. (B2 and C2) W07G4.5::GFP forms a large number of aggregates in
epg-7 mutant embryos. Simultaneously depleting
rheb-1 activity has no effect on the expression of aggregates. (D2-G2) C17E4.2::GFP accumulates into many aggregates in the intestine in
epg-7 mutants (D2 and E2), which disappear after starvation treatment (F2 and G2). (H2-K2) C17E4.2::GFP aggregates in the intestine in
atg-3 mutant larvae are not affected after starvation treatment. (D2, F2, H2, and J2) DIC images of the embryos shown in E2, G2, I2, and K2, respectively. Bars: (A-E and I-C2) 10 um; (insets in M-Q) 5 um; (F-H) 10 um; (D2-K2) 10 um.