Hyperosmotic Stress Induces a Specific Pattern for Stress Granule Formation in Human-Induced Pluripotent Stem Cells

Figure 6

Downregulation of NaCl treatment alters tubulin expression and tubulin network of hiPSCs/IMR90-1. (a) Cells were treated with NaCl for 1 h and then lysed with RIPA buffer. The lysates were separated with SDS-PAGE, transferred onto PVDF membranes, and probed with anti-α-tubulin, anti-β-tubulin, and anti-β-actin antibodies. A representative immunoblot analysis in IMR90/iPSCs and SH-Y5Y cells is shown. (b) Intensity of each band of the immunoblot was measured by the NIH ImageJ program, and the ratios of tubulin and β-actin in each treatment was calculated. (c, d) Representative fluorescence microscopy images showing nontreated hiPSCs/IMR90-1 and SH-SY5Y, cells treated with 200 and 400 mM of sodium chloride stained with the robust SG marker (G3BP (green)), β-tubulin (c) and α-tubulin (d) (red), the largest of the cytoskeletal polymers forming microtubules. Nucleus is stained in blue (Hoechst). Insets show magnified views of SGs and microtubule filaments. Scale bar indicates 5 μm. ns: not significant compared to NT; value less than 0.01 compared to NT.

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