Fig. 5

STAT3 regulates Par3 expression. a Invasion assay. JHOC5 cells were transfected with the STAT3 siRNA (siSTAT3) or control siRNA (siControl). The transfected cells were placed in a Matrigel-coated Boyden chamber 48 h after transfection, and allowed to invade for 24 h. Matrigel membranes were observed with an optical microscope. Scale bar indicates 200 μm. b Wound healing assay. JHOC5 cells were transfected with STAT3 siRNA (siSTAT3) or control siRNA (siControl), seeded onto 6-well culture plates, and grown as a monolayer for 48–60 h until 100% confluent. A scratch assay was then performed. Images of the same area of the wound were taken after 14 h to measure the width of the wound using fluorescence microscope. Data is representative of three independent experiments. c Cell proliferation assay. To analyze the effect of STAT3 knockdown on cell proliferation, 5000 cells were seeded onto 96-well plates 48 h after siRNA transfection. A Cell Counting Kit-8 (Sigma Aldrich) was used to examine proliferation. Data are the mean (±SEM) of four wells. The data shown is representative of three independent experiments. d Effect of STAT3 inhibition on JHOC5 cell proliferation. 5000 cells were seeded onto 96-well plates and incubated for 24 h. Then, they were grown with or without a STAT3 inhibitor (S3I-201) at the doses indicated. Cell proliferation was determined after 24, 48, and 72 h, using a Cell Counting Kit-8 (Sigma Aldrich). Data are the mean (±SEM) of four wells. Showing data is representative of two independent experiments. e Effect of STAT3 inhibition on Par3 expression. JHOC5 cells were treated with or without S3I-201 (100 μM) for 48 h. Cells were lysed and then phospho-STAT3, total STAT3, Par3, and α-Tubulin protein levels were detected by western blotting. f Immunofluorescent analysis of pSTAT3 expression. JHOC5 cells were grown on coverslips and then fixed and stained with anti-phospho-STAT3 and DAPI. Scale bar indicates 10 μm