Fig. 2

Overexpression of HIF-1α in kidney progenitor aggregates. A The mCherry signals in 3D kidney aggregates which were transfected with m-Cherry plasmid or mCherry-HIF-1α overexpressing plasmid for 72 h. B The mCherry signals in 2D kidney progenitor cells with the same treatment as above. C The design for mCherry-HIF-1α overexpressing plasmid. D The transfection efficiency for mCherry positive cells relative to total cells (DAPI) in 2D kidney organoid cells after 72 h transfection. E HIF-1α overexpressing organoids or DMOG treated organoids displayed a higher expression of HIF-1α in immune-fluorescence staining (HIF-1α, red), (Nephrin, yellow), (LTL, green) and (DAPI, blue). Scale bar, 50 μm. F The fluorescent signals for one HIF-1α overexpressing (HIF-1α, red) kidney organoid (LTL, green) with endothelial cells (CD31, yellow) and one normal kidney organoid. G the schematic for HIF-1α overexpression plasmid. H the quantification of immunoreactive signals for HIF-1α. Each dot represents the percentage positive area of immunofluorescent signal in each kidney organoid (n = 4–6 per group, **P < 0.01 and ****P < 0.0001 by unpaired t test). I HIF-1α relative mRNA expression. (n = 5–7 per group, *P < 0.05 and **P < 0.01 by unpaired t test)