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

Figure 6. MAGEA6 mutations and expression variation manipulate autophagy to promote tumor progression at different stages.

(A) Tumor volume plot (50 d after injection) and Kaplan-Meier survival plot (B) of xenograft assay using HPDE-iKRAS cells expressing GFP (Vec) or MAGEA6 (five mice, two injections each, N = 10). P value was calculated by two-tailed unpaired t-test in (A) and log-rank test in (B). (C) (Top) Tumor volume plot (100 d after injection) of xenograft assay using BxPC-3 cells transduced with MAGEA6 shRNAs (#4 and #6) and non-targeting shRNA (NT). P value was calculated by two-tailed unpaired t-test (three mice, two injections each, N = 6). (Bottom) Representative photos and quantification of LC3B immunohistochemical staining in the xenograft tumor samples. P value was calculated by two-tailed unpaired t-test (>150 cells analyzed per cohort). (D) Tumor volume plot (100 d after injection) of xenograft assay using BxPC-3 cells transduced with MAGEA6 (#4), ATG7, VPS34 shRNAs and non-targeting shRNA (NT). P value was calculated by two-tailed unpaired t-test (nine mice, one injection each, N = 9). (E) Patient survival analysis of low and high MAGEA6 expression in stage I (top), stage II (middle), and stage III (bottom) lung cancer. Number of patients analyzed per cohort is shown (Győrffy et al., 2013). P value was calculated by log-rank test.

Figure 6.

Figure 6—figure supplement 1. MAGEA6 expression analysis in PDAC cell line panel.

Figure 6—figure supplement 1.

qRT-PCR of endogenous MAGEA6 expression (A) and immunoblot analysis of endogenous p53 (B) in HPDE and six other PDAC cell lines.
Figure 6—figure supplement 2. Validation of MAGEA6, ATG7 and VPS34 knockdown in BxPC3 cells.

Figure 6—figure supplement 2.

(A) qRT-PCR analysis of MAGEA6 expression of the injected BxPC-3 cells in Figure 6C. P value was calculated by two-tailed unpaired t-test. (B) qRT-PCR analysis of MAGEA6, ATG7 and VPS34 expression of the injected BxPC-3 cells in Figure 6D. P value was calculated by two-tailed unpaired t-test.
Figure 6—figure supplement 3. Gene and protein expression analysis of MAGEA6 variants identified from ICGC lung cancer patients.

Figure 6—figure supplement 3.

Immunoblot (A) and qRT-PCR analysis (B) of MAGEA6 variants identified from the ICGC lung cancer database. MAGEA6 variant expression levels are normalized to those of HEK293T cells transfected with GFP (Vec). MAGEA6 variants with low protein expression are in red.