Figure 6.
Validation of processing defects in other giantin mutant RPE1 lines. (A) Immunoblot of lystates from WT and giantin exon 13 KO RPE1 clones probed for giantin (polyclonal against full-length protein) and tubulin (housekeeping). (B) Widefield maximum-projection images of WT and giantin KO RPE1 cell clones immunolabeled for the N-terminus (N-term) of, and full-length (FL) giantin; ER exit sites (Sec31); and the Golgi (GRASP65) as indicated. Scale bar = 10 µm. (C) Immunoblots of secretion assays performed on WT and giantin KO clones stably expressing GFP-COL1A1 and untransfected giantin KO cells (unTF). Blots show the medium (M) and lysate (L) fractions of each cell population immunoblotted for GFP, SBP, and pro-α1(I) N-propeptide as indicated. (D) Maximum-projection widefield images of GFP signal in giantin KO clones stably expressing GFP-COL1A1 and immunolabeled for pro-α1(I) N-propeptide (magenta). Scale bar = 10 µm.

Validation of processing defects in other giantin mutant RPE1 lines. (A) Immunoblot of lystates from WT and giantin exon 13 KO RPE1 clones probed for giantin (polyclonal against full-length protein) and tubulin (housekeeping). (B) Widefield maximum-projection images of WT and giantin KO RPE1 cell clones immunolabeled for the N-terminus (N-term) of, and full-length (FL) giantin; ER exit sites (Sec31); and the Golgi (GRASP65) as indicated. Scale bar = 10 µm. (C) Immunoblots of secretion assays performed on WT and giantin KO clones stably expressing GFP-COL1A1 and untransfected giantin KO cells (unTF). Blots show the medium (M) and lysate (L) fractions of each cell population immunoblotted for GFP, SBP, and pro-α1(I) N-propeptide as indicated. (D) Maximum-projection widefield images of GFP signal in giantin KO clones stably expressing GFP-COL1A1 and immunolabeled for pro-α1(I) N-propeptide (magenta). Scale bar = 10 µm.

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