ILK knockdown inhibited cell migration and invasion.

<p>(<b>A</b>) Cells from BEL7402 and HLE ILK knockdown stable clones were grown to confluence and a wound was created. Closure of the wound was monitored and captured 16 hours after the wound was made. (<b>B</b>) BEL7402 and HLE ILK knockdown stable clones were seeded onto migration chambers in triplicate and were allowed to migrate for 24 hours. Cells migrated through the membrane were fixed and visualized by crystal violet staining. (<b>C</b>) ILK knockdown stable clones were seeded onto matrigel-coated invasion chamber. BEL7402 was allowed to migrate for 72 hours while HLE required only 24 hours to invade. Cells were fixed, stained and scored. *<i>P</i><0.05 and **<i>P</i><0.001 were regarded as statistically significant.</p

ILK overexpression enhanced PLC cell growth and motility.

<p>(<b>A</b>) FLAG-tagged ILK was transduced into PLC cells and stable clone of ILK was established. Western blot analysis confirmed stable FLAG-ILK expression in PLC cells but not in vector control clone. (<b>B</b>) PLC/vector and PLC/FLAG-ILK cells were counted in triplicates for 8 consecutive days. (<b>C</b>) PLC ILK overexpressing cells were subjected to migration assay. Cells were seeded in triplicates and allowed to migrate for 16 hours. Migrated cells were fixed and stained by crystal violet. (<b>D</b>) PLC and HEK293T cells overexpressing ILK were collected for western blot analysis to study the phosphorylation of Akt and GSK3β. Expression of β-actin was included as an internal loading control. *<i>P</i><0.05 and **<i>P</i><0.001 were regarded as statistically significant.</p

Knockdown of ILK in HCC cell lines impaired HCC cell proliferation and anchorage independent growth.

<p>(<b>A</b>) One sh-non-target control clone (NT) and two ILK knockdown stable clones were established in BEL7402 (E5, E6) and HLE (E5, E7). (<b>B</b>) shILK sequence E6 showed a higher knockdown efficiency than E5 in BEL7402, while both E5 and E7 showed similar knockdown effect in HLE. (<b>C and D</b>) Numbers of cells in BEL7402 and HLE ILK knockdown stable clones were counted in triplicates for 5 to 8 consecutive days. Growth curves were plotted to reveal the growth pattern of each cell line. (<b>E</b>) Cells from BEL7402 ILK knockdown stable clones were grown in soft agar for 1 month. Colonies were formed from single cells and the number of colonies with diameter >60 µm were counted under microscope. (<b>F</b>) Colonies formed from HLE ILK knockdown stable clones were also counted and plotted. Representative pictures of colonies formed from each knockdown stable clone were shown. Asterisk (*) indicates a <i>P</i>-value <0.05 with significant difference between NT control and ILK knockdown clones (E5, E6 and E7).</p

ILK expression was elevated in HCC clinical samples and cell lines.

<p>(<b>A</b>) ILK mRNA expression was examined by qPCR. ILK was overexpressed in 36.9% of the HCC samples. Cases with T/NT ratio >2 were classified as overexpression, while those with T/NT ratio <0.5 were regarded as underexpression. The remaining cases with 0.5< ratio <2 were regarded as no change in ILK expression. Higher ILK expression (<i>P</i> = 0.004) was observed in tumor samples when compared to their non-tumor counterparts. (<b>B</b>) A stepwise increase of ILK expression along HCC tumor stage was observed. (<b>C</b>) ILK expression in a panel of HCC cell lines was analyzed by western blot analysis. MIHA is an immortalized non-tumorigenic liver cell line. Relative ILK expression normalized with corresponding β-actin expression was shown below.</p

ILK knockdown suppressed phosphorylation of Akt and GSK3β.

<p>Insulin was added to cells to stimulate the PKB/Akt signaling pathway. Cell lysates were then collected for western blotting analysis. Expression levels of pAkt, total Akt, pGSK3β, total GSK3β and ILK were shown. Expression of β-actin was included as an internal loading control. The band intensities were determined by densitometry, and the amount of pAkt and pGSK3β was normalized with that of total Akt and GSK3β respectively.</p

Summary of the clinicopathological characteristics of HCC patients.

<p>Summary of the clinicopathological characteristics of HCC patients.</p

Representative Pictures from the Laboratory Section of the Course

<p>Representative Pictures from the Laboratory Section of the Course</p

Discovery-Based Science Education: Functional Genomic Dissection in Drosophila by Undergraduate Researchers

Discovery-Based Science Education: Functional Genomic Dissection in Drosophila by Undergraduate Researcher

Example of the Type of Data Available from the Online Database (http://www.bruinfly.ucla.edu)

<p>Example of the Type of Data Available from the Online Database (<a href="http://www.bruinfly.ucla.edu" target="_blank">http://www.bruinfly.ucla.edu</a>)</p