Image Cytofluorometry for the Quantification of Ploidy and Endoplasmic Reticulum Stress in Cancer Cells.
One of the mechanisms of cancer-associated genomic instability involves a transient phase of polyploidization, in most cases tetraploidization, followed by asymmetric divisions and chromosome loss. Increases in ploidy are consistently accompanied by the activation of an endoplasmic reticulum (ER) stress response, resulting in the translocation of calreticulin to the outer surface of the plasma membrane where it stimulates anticancer immune responses. Conversely, immunoselection leads to a coordinated reduction in ploidy, ER stress, and calreticulin exposure. To simultaneously investigate the ER stress and ploidy, we developed an image cytofluorometric method that allows to measure DNA content, ER stress-associated phosphorylation of eIF2α, and calreticulin exposure at the cell surface. Here, we specify this methodology, which is useful for investigating the correlation between ploidy and ER stress at the single cell level.
Centre de Recherche des Cordeliers, Equipe 11 labellisée Ligue contre le Cancer, INSERM, U1138, Paris, France. laurasenovilla@hotmail.com.
Laura
L
Senovilla
Centre de Recherche des Cordeliers, Equipe 11 labellisée Ligue contre le Cancer, INSERM, U1138, Paris, France.
Yohann
Y
Demont
Centre de Recherche des Cordeliers, Equipe 11 labellisée Ligue contre le Cancer, INSERM, U1138, Paris, France.
Juliette
J
Humeau
Centre de Recherche des Cordeliers, Equipe 11 labellisée Ligue contre le Cancer, INSERM, U1138, Paris, France.
Norma
N
Bloy
Centre de Recherche des Cordeliers, Equipe 11 labellisée Ligue contre le Cancer, INSERM, U1138, Paris, France. kroemer@orange.fr.
Guido
G
Kroemer
D000818:Animals; D002645:Chickens; D059865:Endoplasmic Reticulum Stress / Q000235:genetics / Q000502:physiology*; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism*; D011003:Ploidies; D011817:Rabbits
Calreticulin; ER stress; Image cytofluorometry; Tetraploidy; eIF2α