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Fig. 2 S. aureus supernatant promotes melanoma clustering in vitro. (A) Representative images of ZMEL1-EGFP melanoma cells cultures in the presence of medium alone (Media) or S. aureus USA300 supernatant imaged over 7 days. (B) Plot shows one representative experiment with quantification of melanoma cluster area (µm2) over time. Error bars indicate represent mean±s.e.m. (C,D) Melanoma cluster size (µm2) with S. aureus USA300 supernatant or heat killed bacteria (C) at 7 days or with S. aureus USA300 supernatant plus 500 nM of the ROCK inhibitor Y-27632 (D) at 7 days. Experiments were conducted at least three times. Dots in C and D represent independent replicates. Error bars indicate the mean±s.e.m. P-values were calculated by one-way ANOVA (C) or two-way ANOVA (D). *P<0.05; **P<0.01; ***P<0.001. ns, not significant.