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. 2020 Apr 3;26:111–120. doi: 10.1016/j.jot.2020.03.005

Figure 3.

Figure 3

Effect of Rab27a inhibition on U-MSCs exosomes secretion and yield. (A-B) The protein and mRNA level of Rab27a was decreased in the Rab27a-siRNAsgroup. (B) The expression of exosomal marker (TSG101, CD63 and CD81) in U-MSCs by western blot. (C) The expression of exosomal marker (TSG101, CD63 and CD81) in exosome derived from U-MSCs by western blot. (D) The yield of exosome at static and optimal speed with or without interference. Plots show yield for each method and the mean ± SD of all measurements (∗p < 0.05; ∗∗p < 0.01, n = 3). (E) Therefore, siRNA1 was used for the following interference assay. We cultured U-MSCs transfected with siRNA1 at two RCCS speeds: one completely static (0 rpm/min) and the other at the optimal rotational speed (36 rpm/min). Without mechanical stimulation (0 rpm/min), the exosome yield in the siRNA group was reduced approximately three fold (p<0.01). However, the yield only dropped 1.5-fold in the mechanical environment (p<0.01, Fig. 3E). These results demonstrated that mechanical stimulation can attenuate the reduction in exosome production caused by siRNA interference.