Fig. 2

Knockdown of PTGFRN diminishes cell growth, migration, and invasion in GBM. In U373 cells PTGFRN silenced with either shNT or shPTGFRN, A immunoblot shows protein levels of PTGFRN and β-Actin served as a loading control (required portion of the blot is shown after cropping from the whole blot for both the proteins), B line graph shows the relative cell viability. Representative images show (C) colony number, (D) soft agar colony number, (E) migration, and (F) invasion after silencing PTGFRN in U373, and quantification is shown as bar graphs. (G) Histograms represent the DNA content (stained with PI) in control and PTGFRN silenced cells in U373 and the bar graph represents the quantification of percentage of cells in different phases of cell cycle. (H) Flow cytometry dot plots represent the Annexin-V positive cells in control and PTGFRN silenced cells in U373 and quantification showed as a bar graph, for quantification UR and LR regions of the plot were considered. (I) Immunoblots show the protein levels of PTGFRN, p-ERK, ERK, p-AKT, AKT, p-p70S6, p70S6, p-4EBP1, and 4EBP1 after silencing PTGFRN in U373 and T98G. β-Actin is used as a loading control in western blotting (the required portion of the blot is shown after cropping from the whole blot for all the proteins). The quantification for each blot is given below the blot. The significance was tested using the Student’s t-test and the symbols are indicated as follows: (ns) not significant; (*) p ≤ 0.05; (**) p ≤ 0.01 and (***) p ≤ 0.001