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. 1997 Aug 19;94(17):8970–8975. doi: 10.1073/pnas.94.17.8970

Figure 1.

Figure 1

(A) Northern blot analysis of human MECL-1 expression in T2 cells and transfectants TMP16 (T2 + MECL-1) and T27MP15 (T2 + MECL-1 + LMP2/7). (Upper) Autoradiography of MECL-1 Northern blot, the 1.2-kb RNA marker band is indicated. (Lower) Photography of ethidium bromide-stained total RNA before blotting; 18S and 28S rRNAs are assigned. (B) 2D IEF/PAGE gels of 20S proteasomes purified from T2 cells (T2), TMP16 cells (T2 + MECL-1), and T27MP15 cells (T2 + LMP2 + LMP7 + MECL-1). The subunits LMP2, δ, MECL-1, Z, and LMP7 were assigned according to their migratory position as previously determined by microsequencing analysis. In addition the identity of human MECL-1 and Z was confirmed by N-terminal microsequencing from a Western blot of 2D gels of T27MP15 proteasomes. The sequences obtained were as follows: TTIAGLVFQD for MECL-1 and TTIAGVVYKD for Z. The MB-1 subunit is not visible because its isoelectric point is quite basic, causing it to migrate out of the IEF gel rod under the applied conditions. A total of 60 μg of proteasome was loaded for each gel; proteins were visualized by Coomassie stain.