Skip to main content
. 2012 Jul;78(14):4835–4847. doi: 10.1128/AEM.00431-12

Fig 7.

Fig 7

Intracellular transient expression of tc toxin genes from Y. entomophaga. Expressed cMyc fusion proteins were visualized with Alexa Fluor 594 goat anti-mouse dye-labeled secondary antibody (Molecular Probes) (B, F, J, N, R, V, and Z, compared to AD [control]). Actin was detected with an expressed actin-enhanced green fluorescent protein (EGFP) fusion protein (A, E, I, M, Q, U, Y, and AC), while nuclei were highlighted with Hoechst 33258 DAPI stain (C, G, K, O, S, W, AA, and AE). Merged fluorescence signals are presented in panels D, H, L, P, T, X, AB, and AF. (A to D) Cells transfected with pRK5myc-Chi1 and pEGFP-actin. These cells show actin blebs (yellow arrows) (A and D) and are localized to the cytoplasm (B and D) (red arrows). (E to H) Cells transfected with pRK5myc-Chi2 and pEGFP-actin. As can be observed, pRK5myc-Chi2 causes actin vacuoles (E and H) (yellow arrows) although to a lesser degree than those observed for RK5myc-A1 (I and L) and RK5myc-B (Q and T). Again, pRK5myc-Chi2 is localized to the cytoplasm (F and H). (I to L) Cells transfected with pRK5myc-A1 and pEGFP-actin. pEGFP-actin in RK5myc-A1 shows vacuoles (I and L) (yellow arrows) and nuclear (K and L) and actin (I and L) condensation (white arrows). The RK5myc-A1 tag shows localization to the cytoplasm (J and L) (red arrows). (M to P) Cells transfected with pRK5myc-A2 and pEGFP-actin. pEGFP-actin in RK5myc-A2 shows vacuoles (M and P) (yellow arrows). The RK5myc-A2 tag shows localization to the cytoplasm (N and P) (red arrows). (Q to T) Cells transfected with pRK5myc-B and pEGFP-actin. pEGFP-actin in RK5myc-B shows vacuoles (Q and T) (yellow arrows). Here the RK5myc-B tag shows localization to the nucleus (R and T) (red arrows) and causes nuclear blebbing (S and T) (white arrow). (U to X) Cells transfected with pRK5myc-C1 and pEGFP-actin. As can be seen, pRK5myc-C1 shows actin blebs (U and X) (yellow arrows) and is localized to the cytoplasm (V and X) (red arrows). (Y to AB) Cells transfected with pRK5myc-C2 and pEGFP-actin. As observed for pRK5myc-C1, pRK5myc-C2 shows actin blebs (Y and AB) (yellow arrows) and is localized to the cytoplasm (Z and AB) (red arrows). (AC to AF) Control cells transfected with pEGFP alone. The control cells show normal actin and nuclear morphology and no cMyc colocalization. Scale bars represent 20 μm.