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. 2022 Jun 27;14(8):e15418. doi: 10.15252/emmm.202115418

Figure 3. Characterisation of TPA‐mediated gene delivery.

Figure 3

  • A
    Quantification of gene expression in HEK293 cells after transduction with 0.5 × 106, 1 × 106 and 2 × 106 TU/cell of RGD4C.TPA, RGD4C.AAVP or non‐targeted (NT) particles carrying Lucia. Luciferase activity was measured at days 1 to 6 post‐transduction and shown as relative light units (RLU). Data shown are representative of three independent experiments (n = 3), and are expressed as mean ± SEM. One‐way ANOVA with Tukey's HSD test was used for data analysis.
  • B
    Representative images of FITC‐labelled TPA and AAVP particle diffusion in Matrigel at 5‐ and 30‐min post‐inoculation (n = 3). Scale bar, 100 μm.
  • C
    Particle diffusion using a vertical trans‐well system partitioned by Matrigel. Samples were collected from the wells below the Matrigel at days 1 to 7, n = 3, technical replicates. Data are expressed as mean ± SEM, independent t‐test was used for data analysis.
  • D
    Comparison of AAVP and TPA internalisation efficiency in HEK293 cells as evaluated by staining of the phage capsid proteins, then quantified by FACS at 2‐ and 4‐h post‐transduction, 0.1 × 106 TU/cell, and normalised to particle length. Data are expressed as mean ± SEM of n = 3 independent experiments, independent t‐test was used for data analysis.
  • E
    Quantification of ITRs in the cytosol and nucleus of HEK293 cells 24‐h post‐transduction with 0.5 × 106 TU/cell of RGD4C.TPA and RGD4C.AAVP, using qPCR with primers specific to AAV‐2 ITRs. Data are representative of n = 3 independent experiments and expressed as mean ± SEM, independent t‐test was used for data analysis.
  • F
    Quantification of Lucia reporter gene expression following transduction of human primary SEBTA003 glioblastoma and G26 glioblastoma‐derived neural stem cells with 1 × 106 TU/cell. Data are expressed as mean ± SEM of n = 3 independent experiments. One‐way ANOVA with Tukey's HSD test was used for data analysis.
  • G
    3D primary GBM001 tumour spheres subjected to transduction by increasing doses of TPA and AAVP encoding Lucia. Experiments were repeated three times and performed in triplicates. Data are shown as mean ± SEM and are representative of one experiment. Independent t‐test was used for data analysis.

Source data are available online for this figure.