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. 2021 Oct 6;40(22):e107958. doi: 10.15252/embj.2021107958

Figure 3. Ice2 is required for ER membrane biogenesis upon activation of Ino2/4.

Figure 3

  1. Sec63‐mNeon images of the cortical ER of WT and Δice2 cells harboring the inducible system (SSY1405, 1603). Cells were untreated or treated with 800 nM estradiol for 6 h.
  2. Classification of peripheral ER structures from cortical sections of cells expressing Sec63‐mNeon and Rtn1‐mCherry as tubules (purple), sheets (green), or tubular clusters (yellow). Tubular clusters are combined with tubules in the final classification, as illustrated by the overlay.
  3. Quantification of peripheral ER structures in WT and ∆ice2 cells harboring the inducible system (SSY1405, 1603) and treated with 800 nM estradiol for the times indicated. Bars are the mean percentage of cell cortex covered by tubules (purple) or sheets (green), n = 3 biological replicates. Upper error bars are s.e.m. for the sum of tubules and sheets, and lower error bars are s.e.m. for sheets. Asterisks indicate statistical significance compared with the corresponding value in WT cells, as judged by a two‐tailed Student’s t‐test assuming equal variance. **P < 0.01; n.s., not significant.
  4. mRNA levels of the Ino2/4 target gene INO1 upon ino2* expression in WT and Δice2 cells harboring the inducible system (SSY1405, 1603) as measured by quantitative real‐time PCR. Data were normalized to untreated WT cells. Mean + s.e.m., n = 3 biological replicates. Asterisks indicate statistical significance compared with the corresponding untreated cells, as judged by a two‐tailed Student’s t‐test assuming equal variance. An exception was the test against the normalized value for WT cells, for which a two‐tailed Student's t‐test with unequal variance was applied. *P < 0.05; **P < 0.01.
  5. Quantification of peripheral ER structures in untreated WT, Δice2, Δopi1, and Δice2 Δopi1 cells (SSY1404, 2356, 2595, 2811). Bars are the mean percentage of cell cortex covered by tubules (purple) or sheets (green), n = 3 biological replicates. Upper error bars are s.e.m. for the sum of tubules and sheets, and lower error bars are s.e.m. for sheets. Asterisks indicate statistical significance compared with the corresponding value in WT cells, as judged by a two‐tailed Student’s t‐test assuming equal variance. **P < 0.01; n.s., not significant.

Source data are available online for this figure.