Immunocell-array for molecular dissection of multiple signaling pathways in mammalian cells.
The knowledge of signaling pathways that are triggered by physiological and pathological conditions or drug treatment is essential for the comprehension of the biological events that regulate cellular responses. Recently novel platforms based on "reverse-phase protein arrays" have proven to be useful in the study of different pathways, but they still lack the possibility to detect events in the complexity of a cellular context. We developed an "immunocell-array" of cells on chip where, upon cell plating, growing, drug treatment, and fixation, by spotting specific antibodies we can detect the localization and state of hundreds of proteins involved in specific signaling pathways. By applying this technology to mammalian cells we analyzed signaling proteins involved in the response to DNA damage and identified a chromatin remodeling pathway following bleomycin treatment. We propose our technology as a new tool for the array-based multiplexed analysis of signaling pathways in drug response screening, for the proteomics of profiling patient cells, and ultimately for the high throughput screening of antibodies for immunofluorescence applications.
Zanardi
Andrea
A
Tethis S.r.l., Via Russoli 3, 20143 Milan, Italy.
Giorgetti
Luca
L
Botrugno
Oronza A
OA
Minucci
Saverio
S
Milani
Paolo
P
Pelicci
Pier Giuseppe
PG
Carbone
Roberta
R
D000818:Animals; D001761:Bleomycin / Q000494:pharmacology; D045744:Cell Line, Tumor; D042002:Chromatin Assembly and Disassembly; D004249:DNA Damage; D006801:Humans; D008544:Melanocytes / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D015398:Signal Transduction; D046888:Tissue Array Analysis / Q000379:methods*
D001761:Bleomycin