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. 2010 Aug 9;190(3):407–425. doi: 10.1083/jcb.200911020

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© 2010 Velichkova et al.

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Figure 4. — mtm controls endolysosome homeostasis. (A’–B’, C’, D’, E’, F’, and G’) mtm is necessary and sufficient to restrict size of acidified endolysosomes in Kc₁₆₇ cells (A’–B’, D’, and F’) and primary hemocytes (C’, E’, and G’). (A’ and B’–C’) Normal early endosomes (A’; anti-Rab5) and enlarged and convoluted late endosomes (B’–C’; GFP:Rab7; green) with mtm RNAi are shown. (D’–G’) Both mtm RNAi and null alleles resulted in enlarged acidified endolysosomes (LysoTracker, red; GFP-LAMP, green) that reverted to normal size with mtm:GFP cDNA (G). (H) Area of individual Rab5, GFP:Rab7, and LysoTracker-stained organelles in Kc₁₆₇ cells. (I–J’) Area of LysoTracker-positive organelles decreased in mCherry:mtm-expressing hemocytes. (K and K’) Ultrastructure of mtm RNAi Kc₁₆₇ cells revealed an increase in secondary lysosomes (shaded pink) and electron-lucent membrane compartments (arrowheads). (L and L’) Examples of secondary lysosomes. (M) Secondary lysosome area shown as the percentage of cell area (means: control, 1.8%; mtm RNAi cell area, 5.0%). (N–O′) Examples of electron-lucent membrane structures. Single membrane bound (N and N’) and double membrane bound, typical of autophagosomes (O and O’). (P) Number of individual electron-lucent compartments (means: WT, 1.4; mtm RNAi cell, 6.8). Proportion of single (75 and 78%) and double (25 and 22%) membrane-bound electron-lucent structures remained unaffected. Error bars indicate SEM.