Fig. 6: Enrichment of crystal containing insect cells using fluorescence-based cell sorting.

a Forward (FSC-A) and side scatter (SSC-A) are no viable parameters for cell sorting due to a high similarity in the scattering behavior of crystal-containing and non-containing infected insect cells. b Evaluation of forward (FSC-A) and side scatter (SSC-A) against the fluorescence of EYFP does not produce a selectable population of crystal-containing cells that significantly differs between crystal-containing and non-containing cell cultures. c A high positive correlation is visible between EYFP production and cytosolic as well as endoplasmic target protein production in infected High Five cells. If the target protein is co-translated into the ER, the double negative population shifts towards a single positive population, indicating plasma membrane disruption, while the ER membranes remain intact. d FACS-based selection of infected cells based on their EYFP fluorescence allows an enrichment of crystal containing cells from the original culture. Straight lines in the lower plot indicate the fraction of crystal containing cells before cell sorting. Fractions are corresponding to the gates shown in the upper panel.