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. 2023 Aug 7;42(18):e111620. doi: 10.15252/embj.2022111620

Figure 3. ADPGK‐AS1 overexpression alters the regulation of mitochondrial metabolism.

Figure 3

  • A
    Expression of ADPGK‐AS1 lncRNA (ΔCT) in ADPGK‐AS1 overexpressing (OE) THP1 cells. n = 4 independent experiments. ***P ≤ 0.001 compared with control.
  • B
    Representative FISH images of ADPGK‐AS1 (red) and nuclei (blue) in ADPGK‐AS1 OE THP1 cells. Scale bar = 25 μm.
  • C, D
    Representative images and quantification of mitochondria by MitoTracker (red) and TOM20 (green) in THP1 control and ADPGK‐AS1 OE cells. Nuclei were stained with DAPI (blue). Scale bar = 200 μm (left), 100 μm (right, magnification).
  • E
    Representative immunoblot for mitochondrial protein expression (TOM20, αb‐crystallin, PCG‐1, MRPL15, MRPL35, cytochrome C) in THP1 control and ADPGK‐AS1 OE cells. n = 4 independent experiments.
  • F
    Ratio of mitochondrial DNA (mtDNA) to genomic DNA (gDNA) content in THP1 control and ADPGK‐AS1 OE cells. n = 3 independent experiments. *P ≤ 0.05 compared with control.
  • G
    Gene expression analysis (fold change) of mitochondrially encoded genes (ND1, ND2, ND4, ATP6, ATP8, CO1, CO2, CyB) after isolation of mtDNA from THP1 control and ADPGK‐AS1 OE cells. n = 3 independent experiments. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001 compared with control.
  • H, I
    ATP production (n = 5) and full oxygen consumption plot (n = 8) by the seahorse assay in THP1 control and ADPGK‐AS1 OE cells. **P ≤ 0.01, ***P ≤ 0.001 compared with control.
  • J
    Representative immunoblot of proteins involved in fission and fusion processes of mitochondria (Mfn1, Opa1, Drp1, pAMPK, AMPK). n = 4 independent experiments.
  • K
    Representative confocal fluorescence images and quantification of mitochondrial shape with the mitochondrial marker TOM20 (yellow) in THP1 control and ADPGK‐AS1 OE cells. Nuclei were stained with DAPI (blue). n = 35 cells, Scale bar = 10 μm (left), 5 μm (magnification, right).
  • L
    Transmission electron microscopy images of mitochondrial shapes in THP1 control and ADPGK‐AS1 OE cells. n = 4 cells, Scale bar = 5 μm (left), 1 μm (right, magnification).

Source data are available online for this figure.