MIXL expression is restricted in normal hematopoietic tissues.
(A) Human immune system multitissue Northern blot (Clontech Laboratories) was hybridized overnight with radiolabeledMIXL cDNA probe as described in “Materials and methods,” and the blot was exposed to x-ray film for 5 days. Arrows denote the 3 bands detected. The autoradiograph of the same blot rehybridized with β-actin is shown in the lower panel. PBL indicates peripheral blood leukocytes. (B) The T, B, or myeloid progenitors (1 × 103, 2.5 × 103, and 9 × 103 cells, respectively) from bone marrow or mature T cells, B cells, monocytes, and red blood cells (2 × 105, 1 × 105, 5 × 104, and 1 × 104 cells, respectively) were obtained for total RNA extraction. The RNA from each fraction was reverse-transcribed with an oligo dT primer. The cDNAs were approximately normalized based on a pilot β-actin amplification reaction. Aliquots of 2 μL of the direct or diluted cDNA pool were used in PCR of 18, 21, 24, 27, or 30 cycles with MIXL-EXP primers or β-actin primers as detailed in “Materials and methods.” The amplification products (341 bp for MIXL and 246 bp for β-actin) were resolved on a 1.5% agarose gel, transferred to Hybond N+ membrane, and probed with radiolabeled MIXL or β-actin cDNAs. The blots were exposed to the Phosphor Screen for 1 hour. The images were converted from the Phosphor Screen. The number of PCR cycles is denoted at the bottom.
