Figure S1.
Generation of CRISPR/Cas9 mutant lines. (A) CRISPR design and resulting mutations in chosen KO clones. Top lines show gene sequence in WT and mutant (KO) clones with encoded amino acid sequence underneath. Purple lines indicate gRNA target sequence, scissors point to predicted Cas9 cut site, red letters in KO sequences indicate mutagenic base pair insertions, and purple letters in WT sequence indicate base pairs deleted in the mutant lines. Green amino acids are mutagenic changes arising after frameshift in KO lines, and * denotes a premature stop codon. (B) Maximum projection widefield images of WT and GMAP210 KO lines immunolabeled as indicated. GMAP210 antibodies were raised against amino acids 14–148 (N-term GMAP210), 159–365 (GMAP210), and 1760–1855 (C-term GMAP210). (C) Western blots of WT and GMAP210 KO cells probed with an antibody targeting amino acids 14–148 or GMAP210 (central well is an unsuccessful clone—X). (D) Maximum projection widefield images of WT and Golgin-160 KO clones labeled with GM130 (green, cis-Golgi) and Golgin-160 (magenta). (D) Western blot analysis of WT and Golgin-160 KO cell lysates probed with Golgin-160 antibodies and tubulin and GAPDH as loading controls. (D and E) Golgin-160 N-terminal and C-terminal antibodies raised against amino acids 1–350 and 1436–1498, respectively. Source data are available for this figure: SourceData FS1.

Generation of CRISPR/Cas9 mutant lines. (A) CRISPR design and resulting mutations in chosen KO clones. Top lines show gene sequence in WT and mutant (KO) clones with encoded amino acid sequence underneath. Purple lines indicate gRNA target sequence, scissors point to predicted Cas9 cut site, red letters in KO sequences indicate mutagenic base pair insertions, and purple letters in WT sequence indicate base pairs deleted in the mutant lines. Green amino acids are mutagenic changes arising after frameshift in KO lines, and * denotes a premature stop codon. (B) Maximum projection widefield images of WT and GMAP210 KO lines immunolabeled as indicated. GMAP210 antibodies were raised against amino acids 14–148 (N-term GMAP210), 159–365 (GMAP210), and 1760–1855 (C-term GMAP210). (C) Western blots of WT and GMAP210 KO cells probed with an antibody targeting amino acids 14–148 or GMAP210 (central well is an unsuccessful clone—X). (D) Maximum projection widefield images of WT and Golgin-160 KO clones labeled with GM130 (green, cis-Golgi) and Golgin-160 (magenta). (D) Western blot analysis of WT and Golgin-160 KO cell lysates probed with Golgin-160 antibodies and tubulin and GAPDH as loading controls. (D and E) Golgin-160 N-terminal and C-terminal antibodies raised against amino acids 1–350 and 1436–1498, respectively. Source data are available for this figure: SourceData FS1.

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