Figure 1.
Identification of Bmh1 and Bmh2 as binding partners of Atg11. (A) BY4741 (negative control, NC) or Atg11-3✕FLAG-TEV-ZZ (Atg11-TAP) yeast cells were subjected to glucose starvation for 1 h. Atg11-TAP protein was purified using anti-Rabbit IgG Dynabeads. The samples were separated by 4–12% SDS-PAGE gel, followed by silver staining. Target bands were subjected to LC-MS/MS analysis. (B) Identification of Bmh1/2 proteins by LC-MS/MS analysis. (C and D) Cells co-expressing HA-Atg11 and Bmh1-3✕FLAG (C) or Bmh2-3✕FLAG (D) were treated with glucose starvation for 1 h. Cell lysates were immunoprecipitated with anti-FLAG agarose beads and analyzed by Western blot using anti-HA antibody. The data are representative of three independent experiments. (E and F) Wild-type (WT), atg1∆, atg9∆, atg17∆, atg19∆, or atg20∆ cells co-expressing HA-Atg11 with Bmh1-3✕FLAG (E) or Bmh2-3✕FLAG (F) were subjected to glucose starvation for 1 h. Cell lysates were immunoprecipitated with anti-FLAG agarose beads and analyzed by Western blot using anti-HA antibody. The data are representative of three independent experiments. (G and I) Cells co-expressing HA-Atg11 with Bmh1-3✕FLAG (G) or Bmh2-3✕FLAG (I) were cultured in nutrient-rich medium, nitrogen-starvation medium (SD-N), glucose-starvation medium (SD-G), or treated with rapamycin (0.2 μg/ml) for 1 h. Cell lysates were immunoprecipitated with anti-FLAG agarose beads and analyzed by Western blot using anti-HA antibody. (H and J) The results from G and I were quantified. Data are presented as means ± SD (n = 3). ***P < 0.001; **P < 0.01; two-tailed Student’s t tests were used. Source data are available for this figure: SourceData F1.

Identification of Bmh1 and Bmh2 as binding partners of Atg11. (A) BY4741 (negative control, NC) or Atg11-3✕FLAG-TEV-ZZ (Atg11-TAP) yeast cells were subjected to glucose starvation for 1 h. Atg11-TAP protein was purified using anti-Rabbit IgG Dynabeads. The samples were separated by 4–12% SDS-PAGE gel, followed by silver staining. Target bands were subjected to LC-MS/MS analysis. (B) Identification of Bmh1/2 proteins by LC-MS/MS analysis. (C and D) Cells co-expressing HA-Atg11 and Bmh1-3✕FLAG (C) or Bmh2-3✕FLAG (D) were treated with glucose starvation for 1 h. Cell lysates were immunoprecipitated with anti-FLAG agarose beads and analyzed by Western blot using anti-HA antibody. The data are representative of three independent experiments. (E and F) Wild-type (WT), atg1∆, atg9∆, atg17∆, atg19∆, or atg20∆ cells co-expressing HA-Atg11 with Bmh1-3✕FLAG (E) or Bmh2-3✕FLAG (F) were subjected to glucose starvation for 1 h. Cell lysates were immunoprecipitated with anti-FLAG agarose beads and analyzed by Western blot using anti-HA antibody. The data are representative of three independent experiments. (G and I) Cells co-expressing HA-Atg11 with Bmh1-3✕FLAG (G) or Bmh2-3✕FLAG (I) were cultured in nutrient-rich medium, nitrogen-starvation medium (SD-N), glucose-starvation medium (SD-G), or treated with rapamycin (0.2 μg/ml) for 1 h. Cell lysates were immunoprecipitated with anti-FLAG agarose beads and analyzed by Western blot using anti-HA antibody. (H and J) The results from G and I were quantified. Data are presented as means ± SD (n = 3). ***P < 0.001; **P < 0.01; two-tailed Student’s t tests were used. Source data are available for this figure: SourceData F1.

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