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. 2019 Dec 31;13(1):151–162. doi: 10.15283/ijsc19004

Fig. 3.

Fig. 3

Different hyaluronan substratum exerted changes in mitochondrial distribution of PDMSCs. Successful clones for lentivector pAS3W-DsRed2-mito after ligation of the vectors pLKOAS3w.puro and pDsRed2-mito. (A) PCR gels showed successful clone band at 800 bp; and sequence alignment of 99% from restriction enzyme cutting sites of NheI at 96 bp and PmeI at 892 bp. (B) Transfection of PDMSCs with pAS3W-DsRed2-mito enabled red fluorescence labelling of mitochondria specifically. Lentivector containing DsRed-mito showed fluorescence punctuated patterns when transfected into PDMSCs, whereas lentivector with only DsRed showed a diffused fluorescence pattern; (C) mitochondrial localization of PDMSCs cultured for 5 days on control surfaces, SHA 0.001 and SHA 5 mg/ml or CHA 5 and 30 μg/cm2. (D) Confocal images of mitochondrial distribution in hyaluronan-supplemented PDMSCs. Red fluorescence represents mitochondria through PDMSCs transfected with expression plasmid pAS3W-DsRed2-mito; blue fluorescence represents nucleus through DAPI staining. Scale bar: 100 μm. SHA: medium supplemented hyaluronan, CHA: coated hyaluronan, DAPI: 4’,6-diamidino-2-phenylindole.