Skip to main content
. 2014 Apr 24;124(6):2696–2708. doi: 10.1172/JCI72171

Figure 4. p110α mediates invasion and metastasis.

Figure 4

(A) qPCR analysis of the mRNA levels of genes in the PI3K pathway. Values were normalized on the basis of ribosomal protein L32 mRNA levels and are expressed as ratios (393P_ZEB1 to 393P_vector; 344SQ_miR-200 to 344SQ_vector). (B) Western blot analysis of KP cells. (C) qPCR assays (bar graph) and Western blot analysis (gels) of 344SQ_Pik3ca shRNA cells (shRNA #1 and #2) and 344SQ_scr shRNA cells (Scr). Controls include 344SQ_scr shRNA cells and blotting for actin. qPCR values were normalized on the basis of ribosomal protein L32 mRNA levels and represent the mean ± SD from triplicate samples. (D) Density of 344SQ_Pik3ca shRNA cells and 344SQ_scr shRNA cells in monolayer culture was measured by MTT assay at the indicated time points. Values represent the mean ± SD from triplicate samples. (E) Soft agar colony formation. Colonies were photographed (images) and counted (bar graph) after 14 days of incubation. Original magnification, ×4. Values represent the mean ± SD from triplicate wells. (F) Invasion assays. Cells were photographed (images) and quantified (bar graph). Scale bars: 100 μm. (G) Scatter plots of primary tumor volume (left scatter plot) and numbers of visible lung metastases (right scatter plot) in syngeneic mice injected in the flanks with 344SQ_Pik3ca shRNA (n = 5) or 344SQ_scr cells (n = 5). Each mouse is indicated with a dot. Mean ± SD calculated for each cohort.