Figure S1.
Construction and confirmation of the cell lines used in this study. Gene-targeting strategy, PCR strategy, PCR results, and immunoblotting results are shown for the cell lines used in this study. “P” marks at the top of the panels represent parental lines, whereas numbers (e.g., 4 and 16) indicate line identifications. M, size marker. Asterisks indicate nonspecific bands. (A) mAID-mClover (mAC) tagging to TubG1. PCR amplified a 3.7-kb fragment in the tagged line. Immunoblotting results are shown in Fig. 1 A. (B) Confirmation of TubG2 KO by PCR. The lack of bands derived from exons was confirmed in the KO line. The TubG1 exon and TubG2 UTRs were amplified as positive controls. (C) mAID-mCherry (mACh) or mCherry (mCh) tagging to ch-TOG. PCR amplified a 5.2-kb fragment in the tagged line. Immunoblotting results are shown on the right; the band is shifted upwards with the tag. (D) mAID-mCherry (mACh) tagging to CLASP1. PCR amplified a 5.3-kb fragment in the tagged line. Immunoblotting results are shown on the right; the band is shifted upward with the tag. (E) mAID-mCherry or mCherry tagging to TPX2. PCR amplified a 3.0-kb fragment in the tagged line. Immunoblotting results are shown on the right; the band is shifted upwards with the tag. (F) PCR and immunoblotting confirmation of AKAP450 KO cells. DNA amplification was not observed when a primer targeting an exon was used for the KO line, whereas the hygromycin cassette was amplified with the primers designed at UTRs (this primer set did not amplify very long AKAP450 genes in the parental line). Immunoblotting showed a specific >250-kD band only in the parental line. (G) PCR and immunoblotting confirmation of CAMSAP3 KO. DNA amplification was not observed when a primer targeting an exon was used for the KO line. Immunoblotting showed a specific ∼150-kD band only in the parental line. Puro, puromycin; Neo, neomycin; BSD, blasticidin; Hyg, hygromycin.

Construction and confirmation of the cell lines used in this study. Gene-targeting strategy, PCR strategy, PCR results, and immunoblotting results are shown for the cell lines used in this study. “P” marks at the top of the panels represent parental lines, whereas numbers (e.g., 4 and 16) indicate line identifications. M, size marker. Asterisks indicate nonspecific bands. (A) mAID-mClover (mAC) tagging to TubG1. PCR amplified a 3.7-kb fragment in the tagged line. Immunoblotting results are shown in Fig. 1 A. (B) Confirmation of TubG2 KO by PCR. The lack of bands derived from exons was confirmed in the KO line. The TubG1 exon and TubG2 UTRs were amplified as positive controls. (C) mAID-mCherry (mACh) or mCherry (mCh) tagging to ch-TOG. PCR amplified a 5.2-kb fragment in the tagged line. Immunoblotting results are shown on the right; the band is shifted upwards with the tag. (D) mAID-mCherry (mACh) tagging to CLASP1. PCR amplified a 5.3-kb fragment in the tagged line. Immunoblotting results are shown on the right; the band is shifted upward with the tag. (E) mAID-mCherry or mCherry tagging to TPX2. PCR amplified a 3.0-kb fragment in the tagged line. Immunoblotting results are shown on the right; the band is shifted upwards with the tag. (F) PCR and immunoblotting confirmation of AKAP450 KO cells. DNA amplification was not observed when a primer targeting an exon was used for the KO line, whereas the hygromycin cassette was amplified with the primers designed at UTRs (this primer set did not amplify very long AKAP450 genes in the parental line). Immunoblotting showed a specific >250-kD band only in the parental line. (G) PCR and immunoblotting confirmation of CAMSAP3 KO. DNA amplification was not observed when a primer targeting an exon was used for the KO line. Immunoblotting showed a specific ∼150-kD band only in the parental line. Puro, puromycin; Neo, neomycin; BSD, blasticidin; Hyg, hygromycin.

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