Fig 1: CD40L and 4-1BBL protein content of cells and exosomes(A and B) Exosomes derived from Mel526 cells infected with LOAd viruses or left uninfected were investigated for the presence of CD40L (A) and 4-1BBL (B) with ELISA. Each group was tested in duplicate. Error bars represent standard error and median values are displayed in the graphs.
Fig 2: Surface expression of CD40L and 4-1BBL on exosomes from LOAd-infected cells using flow cytometry(A) Total percentage of CD40L+ and 4-1BBL+ exosomes. (B) Mean fluorescence intensity (MFI) of CD40L and 4-1BBL expression on CD63+ bead-coupled exosomes. ∗p ≤ 0.05, n = 3. Error bars represent standard error and median values are displayed in the graphs.
Fig 3: TMZ-CD40L and 4-1BBL mRNA in infected cells and exosomes (copy number per picogram)(A and B) LOAd-infected or uninfected Mel526 cells and cell-derived exosomes were evaluated for their TMZ-CD40L (A) or 4-1BBL (B) mRNA content using quantitative real-time PCR. Samples were analyzed in triplicates (cells) or duplicates (exosomes). Statistical analyses were done using a Kruskal-Wallis test for non-parametric samples with a Dunn’s multi-comparison test. A p value of =0.05 was considered significant. n = 3.Error bars represent standard error and median values are displayed in the graphs.
Fig 4: Expression of CD40L and 4-1BBL on Mel526 cellsMel526 cells infected with LOAd700 or LOAd703 express CD40L (upper histograms), while cells infected with LOAd703 also present 4-1BBL (lower histograms), as shown with flow cytometry at day 2 post-infection. Gray and black histograms represent isotype control and transgene staining, respectively.
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