Figure 3.

High Expression of Intrinsic MIRO1 by iPSC-MSCs Contributes to Mitochondrial Transfer

(A) The expression of MIRO1 by different types of iPSC-MSCs/ESC-MSCs was detected (Ai and Aii).

(B) (Bi) MIRO1 was measured in iPSC-MSCs transfected with scramble control shRNA (iPSC-MSCs-MIRO1^Sc), MIRO1-shRNA (iPSC-MSCs-MIRO1^Lo), or MIRO1 cDNA plasmid (iPSC-MSCs-MIRO1^Hi). (Bii) The expression of MIRO1 among different groups was evaluated. (Biii) Mitochondrial transfer ratios between GFP-labeled (green) iPSC-MSCs-MIRO1^Sc, iPSC-MSCs-MIRO1^Lo, or iPSC-MSCs-MIRO1^Hi and stressed Celltrace-labeled NMCs (violet) were measured by FACS. We gated out violet+ NMCs and separated them into a violet+/GFP + population with mitochondria transfer (MITO-GFP⁺-NMCs) and a violet+/GFP− population without mitochondria transfer (MITO-GFP⁻-NMCs) to calculate the transfer ratio (violet+/GFP+ cells divided by the total violet+ cells).

(C) (Ci) Western blot analysis for expression of GFP and human COX-4 in the mitochondrial-transferred NMCs that were isolated after co-culture with iPSC-MSCs-MIRO1^Sc, iPSC-MSCs-MIRO1^Lo, or iPSC-MSCs-MIRO1^Hi. The expression of GFP (Cii) and human COX-4 (Ciii) among the different groups was evaluated.

Data represent the means ± SD of three independent experiments. ^∗p < 0.01.