Figure 3.

CCP1 is the sole peptidase that generates ΔC2-α-tubulin in TTLΔ cells. (A,B) siRNA-mediated knock down of CCP1 in TTLΔ cells, followed by detection of ΔC2-α-tubulin with immunofluorescence (A) and immunoblotting (B). In merged images in (A), ∆C2-α-tubulin is shown in magenta, total α-tubulin (DM1A staining) in yellow, and DNA (DAPI staining) in cyan. Scale bars in (A), 50 µm. (C) Detection of CRISPR/Cas9-induced INDELs in the CCP1 locus in TTL∆ CCP1∆ line. Expected premature stop codons were indicated by underlines. (D,E) Detection of ∆C2-α-tubulin in the TTL∆ CCP1∆ line by immunoblotting in cell lysates (D) and immunofluorescence (E). In merged images in (E), ∆Y- or ∆C2-α-tubulin is shown in magenta, total α-tubulin (DM1A staining) in yellow, and DNA (DAPI staining) in cyan. Scale bars in (E), 5 µm.