Ice2 promotes ER membrane biogenesis in yeast by inhibiting the conserved lipin phosphatase complex

A–C

Western blots of FLAG, HA, and Dpm1 from cell lysates or anti‐FLAG immunoprecipitates of WT or ∆nem1 cells containing Spo7‐FLAG, Nem1‐FLAG, or Ice2‐HA as indicated (SSY122, 2421, 3183, 3184, 3195, 3196, 3197). The asterisk marks the position of the light chain from the antibody used for immunoprecipitation. The diamond marks a protein that is non‐specifically precipitated by the anti‐FLAG antibody. IP, immunoprecipitate.

D

Western blot of HA from total cell lysates or biotin pulldowns of cells containing Ice2‐TurboID, Pah1‐HA, and deletions of NEM1 or SPO7 as indicated (SSY2978, 2979, 3117, 3118).

E

Images of cells with endogenously tagged Ice2‐mScarlet and Spo7‐mNeon or Nem1‐mNeon (SSY3244, 3245). Arrows indicate foci containing both Ice2 and either Spo7 or Nem1.

F

Quantification of Spo7 and Nem1 foci in WT and ∆ice2 cells (SSY2916, 3238, 2917, 3239). Mean + s.e.m., n = 3 biological replicates. Asterisks indicate statistical significance compared with the respective WT cells, as judged by a two‐tailed Student’s t‐test assuming equal variance. **P < 0.01.

Figure 7. Ice2 interacts and co‐localizes with the Nem1‐Spo7 complex.