Comparison of hierarchical unsupervised clustering using ROME, RELION, SPIDER and EMAN2.

(A) Unsupervised classification of a MAP2D-generated class into 30 sub-classes using the GTM algorithm in ROME. The red box marks the side view projection of the 11-fold inflammasome complex. The green box marks the side view projection of the 10-fold inflammasome complex, whose length is smaller than that of the others. The yellow box marks the side view projection of the 12-fold inflammasome complex, whose length is larger than that of the others. (B) Unsupervised classification of the same MAP2D-generated class into 30 sub-classes in RELION. The major class exhibits the side view projections of 11-fold inflammasome complex, whereas all the other classes present ‘junk’ features. (C) Unsupervised K-means clustering of the same MAP2D-generated class into 30 sub-classes in SPIDER. The green boxes highlight the side views of the 10-fold inflammasome complex, whose length is smaller than others. The yellow boxes label the side view projections of the 12-fold inflammasome complex, whose length is larger than that of the others. (D) Unsupervised K-means clustering of the same MAP2D-generated class into 30 sub-classes in EMAN2. The yellow boxes label the side view projections of the 12-fold inflammasome complex, whose length is longer than that of the others. The green boxes label the side view projections of 10-fold inflammasome complex, whose length is shorter than that of the others.