Fig. 7

Functional siRNA delivery mediated by G5-cRGD leads to effective anti-proliferative effects in primary SSCs. a Appearance of cell colonies on day 5 after transfection with G5-cRGD-siRNA complexes. Lipo2000 was used as a control. Scale bars: 100 μm. b The mean number of colonies (a cluster consists of more than 5 cells) per square centimeter as shown in (a). c The EdU proliferation assay was performed 7 days after transfection with scrambled siRNA or Cdk1+Cdk2 siRNA by G5-cRGD in primary SSCs. Red: CDH1 used to stain germ cell; green: EdU-positive cell; blue: Hoechst 33342 used to stain nuclei. Scale bars: 50 μm. d Ratios of EdU-positive germ cells to CDH1-positive cells. e Flow cytometry analysis of cell cycle of CDH1⁺ cells was performed 7 days after transfection with scrambled siRNA or Cdk1+Cdk2 siRNA by G5-cRGD. f Quantitation of the cell cycle of the Mock, NC, or KD-(Cdk1+Cdk2) group cells as shown in (e). TUNEL assay was presented by (g) fluorescence microscope images and (h) TUNEL-positive cell counting (%). Red: TUNEL-positive cell; green: CDH1 was used to stain germ cell; DAPI used to stain nuclei. Scale bars: 50 μm. Statistical significance was assessed using one-way ANOVA followed by Dunnett’s test. Data are presented as mean ± standard deviation (SD, n = 3). *p < 0.05, **p < 0.01. Mock: untreated group; NC: scramble siRNA group; KD-(Cdk1+Cdk2): Cdk1+Cdk2 siRNA group