Fig. 5. Reduced Cry1C binding to G2-M-arrested Sf9 cells. (A,C) Cry1C (1 µg/ml in A and 2.5 µg/ml in C) was incubated with normal and G2-M-arrested cells for 5 or 90 minutes. After washing off the non-bound toxin, total proteins (9 µg/lane) were separated by SDS-PAGE and Cry1C binding was detected by western analysis using anti-Cry1C Ab. (Lane 1) Cry1C marker, 100 ng. (Lane 2) Sf9 cells with no Cry1C in the binding reaction. (Lane 3) Sf9 cells after 5 minutes incubation with Cry1C). (Lane 4) G2-M cells after 5 minutes incubation with Cry1C. (Lane 5) Sf9 cells after 90 minutes incubation with Cry1C. (Lane 6) G2-M cells after 90 minutes incubation with Cry1C. Quantification of band-intensity (indicated below the corresponding lanes in arbitrary units) showed that G2-M cells bound 10 times less Cry1C than the untreated cells. (B) Blot shown in (A) probed with anti-actin Ab as a loading control.