Figure 5.

Northern and western-blot analyses of VfPIP. A, Northern-blot analysis of the VfPIP gene in guard cell protoplasts (GCPs), mesophyll cell protoplasts (MCPs), roots, and etiolated seedlings. Each lane contained an equal amount (25 μg) of total RNA isolated from guard cell protoplasts, mesophyll cell protoplasts, roots, and etiolated seedlings. These hybridized with DIG-labeled specific probes of VfPIP gene. Staining of gel with ethidium bromide is shown as a loading control (rRNA). Experiments were repeated three times on different occasions and gave similar results. B, Western-blot analysis using antibodies raised against the C-terminal region of VfPIP as a probe. Each lane contained an equal amount (100 μg) of protein isolated from guard cell protoplasts, mesophyll cell protoplasts, roots, and etiolated seedlings.