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Figure 6


Fig. 6. Production and secretion of LN5 by uPAR-wt and -KO keratinocytes. (A) Suspended (susp.) wt or uPAR-KO keratinocytes, or uPAR-KO keratinocytes infected with a murine uPAR (muPAR) retrovirus, were seeded on a ColIV-coated surface. After 45 minutes, attached cells were detached with 10 mM EDTA and secreted LN5 was detached from the surface with SDS-sample buffer. As a control, suspended cells were lysed. The samples were immunoblotted with anti-LN5 ({gamma}2 chain) antibody. (Right) The amounts of secreted LN5 ({gamma}2 chain) levels in the western blot were quantitated relative to levels in the suspended wt sample (=100%) ± s.d. The values are expressed as the average of at least three experiments. (B) Suspended wt or uPAR-KO keratinocytes, or uPAR-KO keratinocytes infected with a muPAR retrovirus, were seeded on a ColIV-coated surface. After 45 minutes, attached cells were detached for mRNA isolation and the amount of LN5 ({gamma}2 chain) mRNA and actin (control) measured by semi-quantitative RT-PCR. The histogram represents the increase in LN5 mRNA levels relative to the suspended wt sample (=100%). The values are expressed as the average of at least three independent experiments. Error bars represent the s.d.