Skip to main content
. 2022 Jun 13;14(7):e15608. doi: 10.15252/emmm.202115608

Figure 4. tRNA modification deficiency in fibroblasts derived from patients with ELP4 and ELP6 variants.

Figure 4

  1. High‐performance liquid chromatography (HPLC) coupled to mass spectrometry (MS) used to quantify the Elongator‐dependent tRNA modification 5‐carbamoylmethyluridine (ncm5U) in human fibroblasts. Pseudouridine (Ψ) was used as an internal normalization standard. n = 3 technical repeats per genotype.
  2. HPLC‐MS quantification of Elongator‐dependent tRNA modification 5‐methoxy‐carbonylmethyluridine (mcm5U). Pseudouridine (Ψ) was used as an internal normalization standard. n = 3 technical repeats per genotype.
  3. HPLC‐MS quantification of Elongator‐dependent tRNA modification 5‐methoxycarbonylmethyl‐2‐thiouridine (mcm5s2U). Pseudouridine (Ψ) was used as an internal normalization standard. n = 3 technical repeats per genotype.
  4. HPLC‐MS quantification of Elongator‐independent tRNA modification 1‐methyladenosine (m1A). Pseudouridine (Ψ) was used as an internal normalization standard. n = 3 technical repeats per genotype.
  5. HPLC‐MS quantification of Elongator‐independent tRNA modification 7‐methylguanosine (m7G). Pseudouridine (Ψ) was used as an internal normalization standard. n = 3 technical repeats per genotype.
  6. Differential expression (DE) of peptides in fibroblasts obtained from the individuals with ELP4 and ELP6 variants relative to control (n = 3 technical repeats per genotype). Down‐ (red) and upregulated (blue) peptides are depicted on volcano plots.
  7. A network‐based clustering of enriched pathways in ELP4Y91C/L296I was constructed. Text mining was applied to extract the top terms representing each pathway cluster (left). Network interaction maps are shown (right) with the pathway clusters are marked.
  8. A network‐based clustering of enriched pathways in ELP6L118W/L118W was constructed. Text mining was applied to extract the top terms representing each pathway cluster (left). Network interaction maps are shown (right) and the pathway clusters are marked.

Data information: Statistical analysis for data in (A–E): one‐way ANOVA (α = 0.05) with a Dunnett’s post‐hoc test. Statistically significant differences are indicated (*P ≤ 0.05; **P ≤ 0.01; ns, not significant). Data represent mean ± SEM.