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. 2014 Aug 20;24(1):100–117. doi: 10.1093/hmg/ddu422

Figure 5.

Figure 5.

The phenotype observed in ATXN3shRNA is dependent on α5 integrin subunit levels. (A) Overexpression of ITGA5 in ATXN3shRNA cells (ATXN3shRNA_α5) restored their cellular morphology upon RA treatment. Scale bar: 100 μm. (B) ATXN3shRNA_α5 cells no longer presented detectable actin filament disorganization, as shown by phalloidin staining (green). Nuclei were counterstained with DAPI (purple). Upper panel scale bar: 20 μm. Lower panel scale bar: 5 μm. (C) mRNA levels of the neuronal markers analyzed by qRT-PCR in ATXN3shRNA_α5 cells were normal and comparable to the SCRshRNA_α5 control cells. mRNA levels were normalized to undifferentiated cells and HMBS gene. (D) Normalized ITGA5 levels abolished their increased migration of ATXN3shRNA cells in a wound scratch assay, 24 h after scratching. Bars represent migration rate expressed as a percentage of control and calculated calculated as the proportion of the distance between the borderlines caused by scratching, to the distance that remained cell-free after 24 h. (E and F) Normal cell cycle in ATXN3shRNA_α5 cells. n ≥ 3 independent biological replicates in all experiments. *P < 0.05.