Skip to main content
. 2002 Jan;13(1):336–347. doi: 10.1091/mbc.01-02-0302

Figure 6.

Figure 6

Restoration of ROCKI/Rho-kinase expression and activity upon MEK inhibitor treatment of NRK/ras cells. (A) NRK and NRK/ras cells were maintained in the absence (−) or presence (+) of 50 μM PD098059 or 25 μM U0126 for 48 h. Rho-GTP levels were then analyzed (representative of two experiments). (B) Cells were treated as indicated, and whole cell extracts were immunoblotted using anti-ROCKI, anti-Rho-kinase, and anti-β-actin antibodies (top panels). The same extracts were probed with an antiphospho-cofilin antibody, followed by anti-cofilin antibody, after the membrane was stripped (bottom panels). (C) Rho-kinase mRNA expression in NRK and NRK/ras cells upon treatment with PD098059. Rho-kinase cDNAs were amplified by PCR for 15, 20, or 25 cycles from RNAs of NRK and NRK/ras cells incubated in the presence (+) or in the absence (−) of 50 μM PD098059 for 48 h. As a control, rat GAPDH cDNA was also amplified (top panel). Densitometry analysis of Rho-kinase cDNAs amplified for 20 cycles. Data are representative of three independent experiments (mean ± SD). Relative Rho-kinase mRNA levels, normalized to GAPDH levels, are expressed vs. control NRK, which is arbitrarily set to 100% (bottom panel).