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. 2015 Oct 22;6(10):e1933. doi: 10.1038/cddis.2015.288

Figure 4.

Figure 4

LmjMCA is involved in autophagy. (a) Percentage of autophagosome-containing (cells with a punctuated ATG8 staining) WT (dark) and LmjMCA-deficient (gray; Δmca) cells cultivated in a serum-deprived medium: means±S.D. from four independent experiences. WT cells significantly entered autophagy when cultivated in a serum-deprived medium (***P<001, **P<0.01, compared with t0), whereas no significant difference could be detected between the percentage of WT and Δmca autophagosome-containing cells. (b) Growth curves of WT (uninterrupted black line with diamonds), LmjMCA-deficient mutant cells (Δmca; dotted line with squares), Δmca cells expressing ATG8-GFP (Δmca::ATG8; dotted line with crosses) and cells overexpressing the C-terminal domain of LmjMCA (WT[MCA_Cter] uninterrupted gray line with squares) when cultivated in a serum-deprived medium: means±S.D. from minimum seven independent experiences for WT, Δmca and WT[MCA_Cter] and four independent experiences for Δmca::ATG8. Significant growth differences could be observed for Δmca compared with the WT strain, for WT[MCA_Cter] compared with the WT strain at days 1 and 3, and for Δmca::ATG8 compared with the Δmca strain. (c) Growth curves of WT and LmjMCA-deficient strains in normal conditions with 10 μM wortmannin (WT+FCS+WM, dotted black line with squares and Δmca+FCS+WM, dotted gray line with triangles, respectively), of WT strains cultivated in a serum-deprived medium without (WT−FCS−WM, uninterrupted black line with squares) or with 10 μM wortmannin (WT−FCS+WM, dotted black line with circles) and of LmjMCA-deficient cells cultivated in a serum-deprived medium without (Δmca−FCS−WM, uninterrupted gray line with diamonds) or with 10 μM wortmannin (Δmca−FCS+WM, dotted gray line with squares): means±S.D. from minimum three independent experiences. We can note a significant effect of wortmannin at day 1 for WT cells and at day 4 for Δmca, when cells are cultivated in a serum-deprived medium. (d) RT-qPCR quantification of kmp11 (Kinetoplastid Membrane Protein, used as a control) and Lmjmca mRNA expression, after culture of WT cells in a serum-deprived medium (means±S.D. from minimum three independent experiences). We could note a significant overexpression of Lmjmca at day 3 and 4. Unpaired t-test, *P<0.05, **P<0.01 and ***P<0.001