Fig. 10. Role of MCSP in intercellular adhesion. (A) SCC4 cells were incubated in low calcium medium to inhibit intercellular adhesion (–Ca), then transferred to standard medium for 2 hours (+Ca) in the presence or absence of antibodies to E-cadherin (+Ecad) and MCSP (+MCSP), alone or in combination. Cells were labelled with TRITC-conjugated phalloidin. Scale bar, 50 µm. (B,C) Effect of CD8/MCSP on the cohesiveness of keratinocyte clones growing within intact sheets of cultured epidermis. (B) Keratinocytes expressing CD8/MCSP or CD8 were mixed with an excess of keratinocytes transduced with vector alone and allowed to grow to confluence. Cells were then labelled with anti-CD8 followed by Alexa-488-conjugated anti-mouse antibody. Fluorescent images of CD8-expressing clones growing amongst unlabelled neighbours and phase-contrast images of the same cells were collected using a digital camera connected to a fluorescence inverted microscope. Examples of two CD8/MCSP-expressing and two CD8-expressing clones are shown. Scale bar, 40 µm. (C) Clones were scored for cohesiveness. The mean proportions of cohesive colonies from two independent experiments are shown. Error bars represent standard deviation. The total numbers of clones scored in each assay were 78 (CD8) and 53 (CD8/MCSP).

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