Figure 4.

Caspase recruitment domain (CARD)11 is expressed in human skin and increased levels in cutaneous squamous cell carcinoma (cSCC) are associated with nuclear P65. A: Immunofluorescence using a polyclonal antibody raised against recombinant human CARD11 detects low levels of expression in normal human skin compared with cSCC tumors. Control indicates the omission of the primary antibody in normal human skin, DAPI highlights nuclei. B: Raw signal intensities from bead-array experiments performed previously³⁴ were searched for CARD11 and CARD14 expression in vitro (left graph) and in vivo (right graph). Each graph (1 × CARD11, 3 × CARD14) represents a separate probe on the array. In each case expression of CARD11, but not CARD14, was increased in cSCC (n = 5 in vitro, n = 9 in vivo) compared with NHK or normal skin (n = 5). C: Dual immunofluorescence with polyclonal anti-CARD11 and monoclonal anti-active P65 antibodies detects nuclear P65 in cSCC with high CARD11 expression. Scale bars: 100 um (A and C). ^∗∗P < 0.01, two-sample t-test comparison. NHK, normal human keratinocytes (n = 4).