Figure 2.
Generation and expression of COOH- and NH 2 -terminal–truncated constructs of CSQ. (A) Graphic representation of COOH- and NH2-terminal–truncated CSQ constructs with indication of their intracellular localization after transient transfection in L6 cells. The HA epitope was attached at the COOH terminus of only the shortest COOH-terminal–truncated CSQ construct (CSQ1–100). (B–E and F–I) Immunofluorescence of L6 and HeLa cells, respectively, transiently transfected with the various CSQ truncation forms indicated at the top. (J and K) Anti-HA immunofluorescence and LR White–embedded immunogold labeling of HeLa cells transfected with HA-CSQ1–100. (L–O) Immunofluorescence with anti-HA (L and N) and anti-GFP (M and O) Abs in HeLa cells after cotransfection of HA-CSQ1–100 and GFP/CSQ at either high (L and M) or low (N and O) ratio. (P and Q) Immunofluorescence with anti-HA (P) and anti-CSQ (Q) Abs in HeLa cells after transfection of HA-CSQ1–100 (the same as in N and O), but with CSQ1–350 instead of GFP/CSQ. (R) Anti-CSQ Ab, LR White–embedded immunogold labeling of HeLa cell transfected with CSQ1–350. Bar (for K and R): 0.25 μm.