In Situ Protein Validation of Selected Markers from DZ/LZ Differential Gene Signature in Reactive Lymph Node GCs
(A) Double-marker immunofluorescence analysis of Histone H3 expression in the GC showing bright nuclear expression of H3 (green signal) in Ki-67+ (red signal) proliferating cells within the GC DZ.
(B) Triple-marker combined immunohistochemistry/immunofluorescence analysis of MYC (brown signal) expression in the GC showing that scattered MYC+ cells are detected in CD20+ (cyan and red signals) cells within the Ki-67-low (purple and green signals) area corresponding to the LZ.
(C) Triple-marker combined immunohistochemistry/immunofluorescence analysis of CD44 (cyan and red signals) expression in the GC showing that CD44 expression is more intense in Ki-67-low (brown signal) B cells characterized by a brighter CD20 expression (purple and green signals), corresponding with the LZ.
(D) Triple-marker combined immunohistochemistry/immunofluorescence analysis of CD54/ICAM-1 (cyan and red signals) expression in the GC showing that CD54 signal is more intense in Ki-67-low (brown signal) area corresponding to the LZ, where it marks both CD20+ (purple and green signals) and negative (arrowheads) cells. All the representative microphotographs have a 200× original magnification. Scale bars, 100 micrometers.