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. 2018 Mar 29;19(5):e44722. doi: 10.15252/embr.201744722

Figure 2. The formation of stress granules decreases overtime in IDH4 cells exposed to arsenite despite phosphorylation of eIF2α.

Figure 2

  1. Whole‐cell extracts from IDH4 cells treated daily post‐induction of senescence for 30 min with (AS) or without (UNT) 0.5 mM sodium arsenite were prepared and analyzed by Western blot using antibodies for P‐eIF2α, eIF2α, and G3BP1 (loading control).
  2. Whole‐cell extracts from IDH4 cells treated (AS) or not (UNT) with AS after incorporation of 35S‐methionine were run on SDS–PAGE and visualized by autoradiography or Coomassie Blue staining.
  3. (left) After treatment, cells at different stages of senescence [proliferating (PRO), presenescent (PRE), and senescent (SEN)] were fixed, permeablized, and analyzed by immunofluorescence with antibodies against the SG markers FMRP and G3BP1. DAPI staining was used to visualize the nuclei of the cells. Scale bars, 20 μm. (right) Graph represents the number of SGs in the immunofluorescence images (left panel). The average number of SGs per cell was calculated by normalizing the total number of SGs in each field to the total number of cells. Three random fields were used for each quantification. Data are represented as a mean of three independent experiments ± SE (error bars). *P < 0.05 (Student's t‐test).

Source data are available online for this figure.