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. 2019 Jun 21;20(12):3046. doi: 10.3390/ijms20123046

Figure 2.

Figure 2

PXDN stable knockdown in C4-2 cells decreases cell viability and growth on soft agar. (A) PXDN was stably knocked down in C4-2 prostate cancer cells using 4 different shRNA constructs (1F-05, 1F-06, 1F-07, 1F-08) or non-silencing (NS) construct as a control. (A) Fluorescence microscopy (magnification at 20×) was used to detect GFP to confirm successful transduction. (B) Western blot analysis was performed to confirm successful knockdown of PXDN. Actin was utilized as a loading control. (C) Phase contrast microscopy (magnification at 20×) shows the morphology of PXDN knockdown cells. (D) Cell viability was investigated using MTS or Trypan blue assay. (E) Soft agar colony formation was measured in C4-2 NS (control) and representative C4-2 1F-08 (PXDN knockdown) cells. Values were normalized to C4-2 NS and the mean± SEM of data were obtained from three independent replicate experiments. Statistical analysis was done with GraphPad Prism; (**** p < 0.0001, *** p < 0.001, ** p < 0.01, * p < 0.05).