Time-course Analysis of PACAP-induced Transcriptome and Proteome in PC12 Cells

The PC12 cells (RCB0009) were obtained from the RIKEN Cell Bank (Tsukuba, Japan). Cells were cultivated in RPMI medium 1640 (ATCC Modification, Thermo Fisher Scientific, Grand Island, NY, USA) in a CO₂ incubator (37 °C, 5%) with 5% fetal bovine serum (FBS) (16140063, Gibco, Billings, MT, USA), 10% horse serum (HS) (H1138, SIGMA, St. Louis, MO, USA), and antibiotics (penicillin-streptomycin, P4458, Sigma, St. Louis, MO, USA). Cellmatrix Type IV (Nitta Gelatin Inc., Osaka, Japan) was used to coat the culture dish. PC12 cells cultured in 10 cm sterile culture dishes were incubated for 24 h, and PACAP38 10⁻⁷ M was added post-incubation. PC12 cells after 5, 10, 15, 30, 60, and 120 min post-incubation from PACAP38 addition were used for total RNA extraction for DNA microarray analysis [9 (link),10 (link),11 (link),12 (link)]. Similarly, PC12 cells after 15, 30, 60, and 120 min of incubation from PACAP addition were used for total protein extraction for proteomic analysis [9 (link),10 (link),12 (link)].

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Shibato J., Takenoya F., Yamashita M., Gupta R., Min C.W., Kim S.T., Kimura A., Takasaki I., Hori M., Shioda S, & Rakwal R. (2023). OMICS Analyses Unraveling Related Gene and Protein-Driven Molecular Mechanisms Underlying PACAP 38-Induced Neurite Outgrowth in PC12 Cells. International Journal of Molecular Sciences, 24(4), 4169.

Publication 2023

RPMI medium 1640 FBS H1138 penicillin-streptomycin Cellmatrix Type IV

Antibiotics Cells Dishes Dna microarray Fetal bovine serum Gelatin Horse Microarray analysis Pacap Pacap38 Pc12 cells Penicillin Protein Serum Sterile Streptomycin

Corresponding Organization : Shonan University of Medical Sciences

Other organizations : Hoshi University, Ebara (Japan), Kookmin University, Pusan National University, University of Toyama, Tohoku University, University of Tsukuba

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Variable analysis

independent variables

PACAP38 concentration (10^-7 M)

dependent variables

Total RNA expression for DNA microarray analysis
Total protein expression for proteomic analysis

control variables

Cell line (PC12 cells, RCB0009 from RIKEN Cell Bank)
Culture medium (RPMI 1640 with 5% FBS, 10% HS, and antibiotics)
Culture conditions (37 °C, 5% CO2)
Culture dish coating (Cellmatrix Type IV)
Incubation time points (5, 10, 15, 30, 60, 120 min for RNA; 15, 30, 60, 120 min for protein)

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