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. 2023 Jul 10;42(16):e112446. doi: 10.15252/embj.2022112446

Figure 5. Storage body components bind MTPTs and associate with mitochondria in a local translation‐dependent manner.

Figure 5

  1. Analysis of expression of select MTPTs by RT–qPCR following RNA immunoprecipitation (RIP) in anti‐GFP isolates from NTL‐2::GFP transgenic animals, unc‐119(ed3)III (no GFP expression) and p gst‐4 GFP (used as an additional negative control) counterparts (n = 2 independent experiments, ***P < 0.001; one‐way analysis of variance (ANOVA)).
  2. Representative images showing the localization of NTL‐2/storage bodies relative to mitochondria upon genetic inhibition of either tomm‐20 or akap‐1 (green: NTL‐2, red: TMRE, a mitochondrial membrane potential‐dependent dye; n = 3 independent experiments). Scale bars, 20 μm. Images were acquired using an ×63 objective lens.
  3. Quantification of the distances NTL‐2/storage bodies acquire from mitochondria upon genetic inhibition of either tomm‐20 or akap‐1 (n = 3 independent experiments with at least 45 animals/experiment; ***P < 0.001; one‐way analysis of variance (ANOVA)).
  4. Quantification of the number of NTL‐2/storage bodies upon genetic inhibition of either tomm‐20 or akap‐1 (n = 3 independent experiments with at least 45 animals/experiment; **P < 0.01, ***P < 0.001; one‐way analysis of variance (ANOVA)).
  5. Immunoblot analysis in mitochondria isolates of whole animal extracts showing the protein levels of NTL‐2 contained in the isolate in control conditions and upon genetic inhibition of akap‐1, tomm‐20, mrps‐5 and atp‐3; genetic inhibition of ntl‐2 is used as a control; NTL‐2 protein is detected by an anti‐GFP antibody in transgenic animals that contain the NTL‐2::GFP protein fusion (n = 2 independent experiments).
  6. Quantification of NTL‐2 protein bound to mitochondria under control conditions and upon genetic inhibitions shown in (E); CTC‐1 is used as a loading control for mitochondria (n = 2 independent experiments; *P < 0.05, **P < 0.01, ***P < 0.001; one‐way analysis of variance (ANOVA) followed by Dunnett's test).

Data information: Error bars denote SEM.