FIGURE

Figure 4

ID
ZDB-FIG-200312-10
Publication
Hoelscher et al., 2020 - miR-128a Acts as a Regulator in Cardiac Development by Modulating Differentiation of Cardiac Progenitor Cell Populations
Other Figures
All Figure Page
Back to All Figure Page
Figure 4

Knockdown of miR-128a during in vitro differentiation of murine Isl1-Cre/ROSA26mTmG induced pluripotent stem cells (iPSCs) (iITG-iPSCs). (A) iITG-iPSCs (scale bar: 500 µm) were differentiated using the hanging drop method. Transfections of LNA probes for knockdown of miR-128a (LNA-128) and corresponding controls (LNA-Ctr) were performed after 0.75 weeks (wks) and after 2 wks of differentiation. Analysis was performed on respective timepoints between 1 and 3 wks. Abbreviations: EB: Embryoid bodies; FACS: Flow cytometry; (B) MiR-128a was significantly downregulated by LNA-128 from 1 wk until 3 wks compared to respective LNA-Ctr (n = 8,7, Mann–Whitney tests, 3 assays in triplicates). (C) Isl1-GFP-positive CPCs after 2 wks (upper panel, scale bars: 500 µm) and after 3 wks (lower panel, scale bars: 500 µm) of in vitro differentiation after transfection with either LNA-Ctr or LNA-128. Images are an overlay between phase contrast and fluorescent microscopic pictures. (D) The frequency of Isl1-GFP-positive CPCs after miR-128a knockdown was significantly increased to the double amount compared to LNA-Ctr (n = 9, p = 0.024, t-test; 3 assays in triplicates) after 2 wks. At later timepoints, the Isl1-CPC frequency was not significantly affected by miR-128 knockdown (3 assays in triplicates at 2.25 wks, 2 assays in triplicates at 3 wks). (E) Isl1 expression was tendentially upregulated during iITG-iPSC differentiation after miR-128 knockdown (n = 6, 2 assays in triplicates). (F) Beating frequency (beats per minute, bpm) of differentiated iITG-iPSC EBs was reduced after miR-128 knockdown (2 and 2.25 wks: 3 assays in triplicates, 2.75 (t-test) and 3 wks (Mann–Whitney test): Two assays in triplicates, 4–9 videos per condition, 3 independent observers). Data are represented as means ± SEM. * p ≤ 0.05, ** p ≤ 0.01 and *** p ≤ 0.001.

Expression Data

Expression Detail
Antibody Labeling
Phenotype Data

Phenotype Detail
Acknowledgments
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Int. J. Mol. Sci.