Fig. 5: Intercellular mitochondrial transport and therapeutic potential of MSC-Ob are restored upon culturing in PQQ.

A MSCs were transduced with mito-GFP and co-cultured with MLE12 which were treated with Veh or Rot and stained with CTDR. The % GFP signal was counted in by gating MLE12 cells after 24 h. B, C Similarly, mitoSOX and TMRE staining was done in MLE12 cells after co-culture with MSCs and represented as TMRE fluorescent intensity (FI). D CTDR stained MLE12 cells were co-cultured with mito-GFP transduced MSCs and further stained with MTR. The MTR images were taken from MLE12 cells to determine mitochondrial size distribution after 48 h of co-culture. E Similar to D, with integrated density representing the mitochondrial mass in MLE12 cells by specifically counting the MTR signal in these cells. F MLE-12 cells were stained with CTDR and co-cultured with untagged MSCs for 24 h. The cells were stained with propidium iodide (PI), and flow cytometry analysis was done. MLE12 cells were gated using CTDR, and the PI signal was calculated, which is represented as % MLE12 cell death. G Immunoblots and densitometry analysis (lower panel) of Miro1 and β-actin in cell lysates prepared from various groups of MSCs. H TNT formation between co-cultured MSCs and MLE12 cells was determined by counting the number of physically attached TNTs. The data is represented as no. of TNTs per 10² cells. Mean ± SEM. ****P < 0.001; ***P < 0.005; **P < 0.01; *P < 0.05; ns (non-significant).