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3 protocols using cos 7 cells

1

Fluorescent Labeling of Cellular Structures

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Modified DNA oligonucleotides were purchased from Bioneer (Daejeon, Republic of Korea). AF488 (Alexa Fluor 488 NHS Ester, catalog number: A20000) and Nunc Lab-Tek chambered coverglass (catalog number: 155383PK) were purchased from Thermo Fisher Scientific. CF488A (CF®488A Succinimidyl Ester, catalog number: 92120) and CF660R (CF®660R Succinimidyl Ester, catalog number: 92134) were purchased from Biotium. Cy5 (Cy5 NHS Ester, catalog number: PA15101) was purchased from GE Healthcare Life Sciences. COS-7 cells were purchased from Korean Cell Line Bank. Anti-tubulin antibody (catalog number: ab6160) was purchased from Abcam. Donkey anti-rat IgG antibody (catalog number: 712–005-153) was purchased from Jackson ImmunoResearch Laboratories, Inc. The docking strands were conjugated to the secondary antibodies using Antibody-Oligonucleotide All-in-One Conjugation Kit (catalog number: A-9202-001) purchased from Solulink. Glutaraldehyde (catalog number: G5882), Triton X-100 (catalog number: T9284), Sodium Borohydride (catalog number: 452882-5G), and Bovine Serum Albumin (catalog number: A4919) were purchased from Sigma-Aldrich.
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2

Fluorescently-labeled Antibody Conjugation

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Modified DNA oligonucleotides were purchased from Integrated DNA Technologies. Alexa488 (Alexa Fluor 488 NHS Ester, catalog number: A20000) was purchased from Thermo Fisher Scientific. Cy3 (Cy3 NHS Ester, catalog number: PA13101) and Cy5 (Cy5 NHS Ester, catalog number: PA15101) were purchased from GE Healthcare Life Sciences. COS-7 cells were purchased from Korean Cell Line Bank. Anti-tubulin antibody (catalog number: ab6160) was purchased from Abcam. Anti-Tom20 antibody (sc-11415) was purchased from Santa Cruz Biotechnology, Inc. Donkey anti-rabbit IgG antibody (catalog number: 711-005-152) and donkey anti-rat IgG antibody (catalog number: 712-005-153) were purchased from Jackson ImmunoResearch Laboratories, Inc. Carboxyl latex beads (catalog number: C37281) were purchased from Thermo Fisher Scientific. The docking strands were conjugated to the secondary antibodies using Antibody-Oligonucleotide All-in-One Conjugation Kit (catalog number: A-9202-001) purchased from Solulink. Paraformaldehyde (catalog number: 1.04005.1000) was purchased from Merck. Glutaraldehyde (catalog number: G5882), Triton X-100 (catalog number: T9284), and Bovine Serum Albumin (catalog number: A4919) were purchased from Sigma-Aldrich.
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3

In Vitro Imaging of COS-7 and NS-1 Cells

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For the in vitro imaging, COS-7 cells (#21651, Korean Cell Line Bank) and Neuroscreen-1 cells (NS-1, subclone of PC12 cells; Cellomics) were used. COS-7 cells were maintained in 90% Dulbecco's modified Eagle's medium (DMEM, #10-013-CV, Corning) with 10% heat-inactivated fetal bovine serum (FBS, #10082147, GIBCO, Life technologies) at 37 C and 10% CO 2 . NS-1 cells were provided by T. Meyer (Stanford University) and maintained in 80% Ham's F-12K (Kaighn's) Medium (#21127022, GIBCO, Life technologies) with 15% heat-inactivated horse serum (#26050088, GIBCO, Life technologies) and 5% heat-inactivated FBS (#10082147, GIBCO, Life technologies) at 37 C and 5% CO 2 . All cell lines were confirmed to be free from mycoplasma by using e-Myco Mycoplasma PCR detection kit (ver. 2.0, iNtRON).
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