Deficiency in retrograde sterol transfer leads to large PM invaginations. (A) Internal mCherry-D4H signal in CK-666–treated ltc1Δ cells form deep invaginations colabeled by the PM t-SNARE GFP-Psy1. (B) Correlative TEM and epifluorescence (mCherry-D4H) images of a 300-nm section of mCherry-D4H–expressing ltc1Δ cells treated with CK-666 for 45 min to 1 h. The overlay is shown on the left. Virtual sections of tomographic reconstruction of the regions highlighted by gray boxes are shown on the right. Three examples show (i) invaginations from the PM, (ii) large internal bulging region, and (iii) tube surrounded by ER (blue arrowheads). Yellow arrowheads point to D4H-positive organelles. (C) PM invaginations labeled by sfGFP-D4H in CK-666–treated ltc1Δ mutant are surrounded by ER marked by mCherry-AHDL. An enlargement of the boxed region is shown over three z-stacks on the right. (D) PM invaginations labeled by sfGFP-D4H in CK-666–treated ltc1Δ form at eisosomes, marked by Pil1. In WT cells, Pil1 and D4H do not colocalize. In ltc1Δ cells, Pil1 appears to surround the D4H signal on PM invaginations. Images on the left are maximum projections of 0.4-µm-spaced z-planes. Scale bars, 200 nm in B; 0.5 µm in C, right panel; and 2 µm elsewhere.
Figure 7.

Deficiency in retrograde sterol transfer leads to large PM invaginations. (A) Internal mCherry-D4H signal in CK-666–treated ltc1Δ cells form deep invaginations colabeled by the PM t-SNARE GFP-Psy1. (B) Correlative TEM and epifluorescence (mCherry-D4H) images of a 300-nm section of mCherry-D4H–expressing ltc1Δ cells treated with CK-666 for 45 min to 1 h. The overlay is shown on the left. Virtual sections of tomographic reconstruction of the regions highlighted by gray boxes are shown on the right. Three examples show (i) invaginations from the PM, (ii) large internal bulging region, and (iii) tube surrounded by ER (blue arrowheads). Yellow arrowheads point to D4H-positive organelles. (C) PM invaginations labeled by sfGFP-D4H in CK-666–treated ltc1Δ mutant are surrounded by ER marked by mCherry-AHDL. An enlargement of the boxed region is shown over three z-stacks on the right. (D) PM invaginations labeled by sfGFP-D4H in CK-666–treated ltc1Δ form at eisosomes, marked by Pil1. In WT cells, Pil1 and D4H do not colocalize. In ltc1Δ cells, Pil1 appears to surround the D4H signal on PM invaginations. Images on the left are maximum projections of 0.4-µm-spaced z-planes. Scale bars, 200 nm in B; 0.5 µm in C, right panel; and 2 µm elsewhere.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal