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. 2018 Jul 17;9:1588. doi: 10.3389/fmicb.2018.01588

FIGURE 5.

FIGURE 5

TLR2 signaling mediates the effects of PSA on activation of IL-8 production, and the suppression of proliferation, migration, and invasion of SW620 cells. SW620 colorectal cancer cells were transduced with either nonsense control (shCtrl) or two different TLR2 shRNAs targeting different regions of TLR2 (shTLR2#1 or shTLR2#2), and the transfected cells were selected with puromycin. The cell lysate was obtained from the transduced cells and western blot analysis was performed to analyze the TLR2 protein level; ROD, relative optical density, β-actin was used as the loading control (A). The shCtrl or two different clones of shTLR2-transduced cells were treated with 100 μg/ml PSA for 24 h. The production of IL-8 was assessed by ELISA (B). The shCtrl and shTLR2-transduced cells were treated with 10, 50, or 100 μg/ml PSA for 48 h to determine the cell viability by the MTT assay (C). The shCtrl and shTLR2-transduced cells were treated with 100 μg/ml PSA for 24 h. Relative mRNA expression levels of cell cycle-related genes (CCNB1, CCND1, CDK2, and CDKN1B) were measured (D). Cell migration was observed by a Transwell migration assay (E), cell invasion was also evaluated by a Matrigel-coated Transwell assay (F). Data are shown as the mean ± SD from three independent experiments (∗P < 0.05, ∗∗P < 0.005, ∗∗∗P < 0.0005).