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Apg7pG333A and Apg7pC507A show significant defects in the formation of the Apg12p–Apg5p conjugate. The conjugate in cell lysates expressing HA-tagged Apg12p was detected by Western blotting using αHA antibody as described previously (Mizushima et al. 1998a blue right-pointing triangle). pAPG7myc-314/pRS316: apg7Δ cells expressing c-myc–tagged wild-type Apg7p only; pRS314/pAPG12HA-316: apg7Δ cells expressing HA-tagged Apg12p only; pAPG7myc-314/pAPG12HA-316: strain YIT702 cells; pAPG7G333Amyc-314/pAPG12HA-316: strain YIT7G333A cells; pAPG7C507Amyc-314/pAPG12HA-316: strain YIT7C507A cells.

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