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. 2020 Apr 9;9:e48963. doi: 10.7554/eLife.48963

Figure 5. Overexpression of wild-type MAGEA6, but not mutant MAGEA6, suppresses autophagy in PDAC cell lines.

(A) Immunoblot analysis of autophagy signaling in HPDE-iKRAS cells expressing GFP (Vec) and MAGEA6 variants. (B) Immunofluorescence staining of LC3B puncta in the transduced HPDE-iKRAS cells. Representative photos (left) and statistical analysis (mean ± standard deviation of counted cells, N=~100 per cohort) are shown. *p=0.002; two-tailed unpaired t-test. (C) Immunoblot analysis of autophagy substrate SQSTM1/p62 in the transduced HPDE-iKRAS cells treated with BafA1 for the indicated time points., Immunoblot analysis of (D) autophagy signaling and (E) SQSTM1/p62 accumulation in wild-type MAGEA6 expressing and (F) autophagy signaling in MAGEA6H305fs* expressing HPDE-iKRAS cells under nutrient-deficient conditions. (G) Immunoblot analysis of autophagy signaling in wild-type MAGEA6 expressing cells under prolonged nutrient-deficient conditions.

Figure 5.

Figure 5—figure supplement 1. Immunoblot analysis of autophagy signaling of transduced AsPC-1 and MIA PaCa-2.

Figure 5—figure supplement 1.

Figure 5—figure supplement 2. Immunoblot analysis of the accumulation of autophagy substrate SQSTM/p62 in the transduced HPDE-iKRAS cells under BafA1 for the indicated time points.

Figure 5—figure supplement 2.

Figure 5—figure supplement 3. Immunoblot analysis of autophagy activity in transduced HPDE-iKRAS cells under nutrient-depleted conditions as indicated.

Figure 5—figure supplement 3.