Extended Data Fig. 5: Single cell cloning classifies A431 cells in inducers and non-inducers.
A) qRT-PCR of IFNB1 and MX2 in direct, indirect and alone A431/VCAF2b co-cultures after siRNA KD of E-cadherin. Each dot is a biological replicate (IFNB1 n=4, MX2 n=3). mRNA levels shown were normalized to siRNA scramble control. One sample t test. B) Images show DAPI staining of A431 cells treated with DMSO or 1μM AZD6733. Orange arrows show micronuclei. Images are representative of three independent experiments. Scale bar is 10 μm. C) Micrographs of IF staining of replicated cytoplasmic DNA labelled with BrdU (red), DAPI (blue) and Phalloidin (cyan) in A431 inducers and non-inducers. White arrows highlight presence of replicated DNA in the cytoplasm of Inducer cells. Images are representative of two independent experiments. Scale bar is 20μm. D) qRT-PCR comparing MX2 expression in direct co-cultures (Touch) of ‘inducer’ and ‘non-inducer’ A431 with VCAF2b. On the right, enlarged graph showing qRT-PCR comparing MX2 expression between direct (Touch) and indirect (No Touch) co-cultures of non-inducers A431 with VCAF2b. Each dot is a biological replicate (inducers n=5, non-inducers n=3). mRNA levels were normalized against two housekeeping genes. E) FACs plot showing equivalent numbers between inducers and non-inducers of VCAF2b cells with A431 cytoplasm fragments as seen in the GFP channel. Representative FACS plot from two independent experiments. For all graphs; each dot is a biological replicate, number of independent experiments is shown in the figure, represented is mean with SD. See also Statistical Source Data Extended Data Fig.5.
