Differential expression of adseverin and gelsolin spots in Lin^–Sca⁺Kit^– and Lin^–Sca⁺Kit⁺ cells from murine bone marrow. (A) Protein spots yielding tryptic mass fingerprints (using MALDI mass spectrometry) characteristic of adseverin and gelsolin are highlighted in white (for adseverin) and orange (for gelsolin). (B) Areas corresponding to the boxed regions in A are shown in 3D pseudocolor format. (C) Histograms of the normalized volumes of the spot chains Lin^–Sca⁺Kit⁺ (▦) and Lin^–Sca⁺Kit^– (▪) (from left to right: 1-7 for adseverin, shown in white; 1 and 2 for gelsolin, shown in black) for gelsolin (i) and adseverin (ii). Normalized volume was calculated on the basis of spots present in all gels. (Di) The relative expression levels of adseverin and gelsolin mRNA in primitive hematopoietic cells were assessed by semiquantitative PCR using GAPDH as a loading control. Transcripts were amplified and visualized by agarose gel electrophoresis. (Dii) Amalgamated results of RT-PCR reactions from a dilution series of 10%, 20%, 50%, and 150% of cDNA for each primer combination. The data illustrate the semiquantitative nature of the RT-PCR. Error bars indicate SEM of 3 observations.