Figure 4.
Erg6 and Pln1 exhibit LD targeting preferences in human HeLa cells independent of AGR stress.(A) Cartoon of HeLa cell lipid treatments with BSA-conjugated OA or methyl-β-cyclodextrin-conjugated CHOL, generating TG-rich or SE-rich LDs. (B) Yeast expressing Erg6-DsRed2 and stained for LDs with MDH, then imaged either in log-phase growth (top) or following 4 h AGR. (C) HeLa cells were transfected for 24 h with Erg6-eGFP and then incubated with 400 µM of OA or 250 µM of methyl-β-cyclodextrin conjugated CHOL for 24 h. Scale bars indicated. LDs stained with LipidTox or MDH. (D) HeLa cells transfected for 24 h with Erg6-DsRed2 and then incubated with 400 µM of OA or 250 µM of methyl-β-cyclodextrin conjugated CHOL for 24 h. (E) Analysis of the quantified LD-to-ER targeting ratios of Erg6-eGFP or Erg6-DsRed2 after induction of TG or SE-rich LDs. Mean ± SEM n = 340–989 LDs from 19 to 28 cells. Triplicate experiments were conducted. (F) HeLa cells were transfected for 24 h with Pln1-eGFP and then incubated with 400 µM of OA or 250 µM of methyl-β-cyclodextrin conjugated CHOL for 24 h. Scale bars indicated. LDs stained with LipidTox. (G) Analysis of the quantified LD/cytosol targeting ratios of Pln1-eGFP after induction of TG or SE-rich LDs. Mean ± SEM, n = 19–21 cells. n = 893 for TG-rich LDs, 268 for SE-rich LDs from (respectively) 14 and 12 cells. Triplicate experiments were conducted. ****, P < 0.00001. All scale bars are indicated.

Erg6 and Pln1 exhibit LD targeting preferences in human HeLa cells independent of AGR stress . (A) Cartoon of HeLa cell lipid treatments with BSA-conjugated OA or methyl-β-cyclodextrin-conjugated CHOL, generating TG-rich or SE-rich LDs. (B) Yeast expressing Erg6-DsRed2 and stained for LDs with MDH, then imaged either in log-phase growth (top) or following 4 h AGR. (C) HeLa cells were transfected for 24 h with Erg6-eGFP and then incubated with 400 µM of OA or 250 µM of methyl-β-cyclodextrin conjugated CHOL for 24 h. Scale bars indicated. LDs stained with LipidTox or MDH. (D) HeLa cells transfected for 24 h with Erg6-DsRed2 and then incubated with 400 µM of OA or 250 µM of methyl-β-cyclodextrin conjugated CHOL for 24 h. (E) Analysis of the quantified LD-to-ER targeting ratios of Erg6-eGFP or Erg6-DsRed2 after induction of TG or SE-rich LDs. Mean ± SEM n = 340–989 LDs from 19 to 28 cells. Triplicate experiments were conducted. (F) HeLa cells were transfected for 24 h with Pln1-eGFP and then incubated with 400 µM of OA or 250 µM of methyl-β-cyclodextrin conjugated CHOL for 24 h. Scale bars indicated. LDs stained with LipidTox. (G) Analysis of the quantified LD/cytosol targeting ratios of Pln1-eGFP after induction of TG or SE-rich LDs. Mean ± SEM, n = 19–21 cells. n = 893 for TG-rich LDs, 268 for SE-rich LDs from (respectively) 14 and 12 cells. Triplicate experiments were conducted. ****, P < 0.00001. All scale bars are indicated.

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