Figure 1.

Detection of the splice variant 2 of Siglec-11 in human brain tissue. A, Gene transcript analysis of three different human brain tissues samples (H. brain 1, 2, and 3) by RT-PCR. Amplification with three different pairs of primers (primer pair 1, 2, and 3) from distinct exons resulted in PCR products of 103 bp, 352 bp, and 294 bp, respectively. All three PCR products were sequenced and showed gene sequence of Siglec-11 splice variant 2. Control, RNA of tissue sample 1 without reverse transcription. M. brain, cDNA from mouse brain tissue. Marker, 100 bp ladder. B, Western blot analysis of protein lysates from distinct human brain tissue samples (brain 1, 2, and 3), normal 293 cells (293), and 293 cells transduced with the Siglec-11 vector (293-Siglec-11). Siglec-11 protein was detected in the brain tissue and the positive control (293-Siglec-11). Antibody directed against β-actin, which labeled a 42 kDa protein, was applied as control. C, Immunofluorescence analysis of Siglec-11 in human brain tissue samples. Brain tissue sections were double immunolabeled with antibodies directed against Siglec-11 and Iba1. Siglec-11 expression was detected in microglial cells by confocal laser microscopy. Background staining was determined by control antibodies (control). Scale bars, 15 μm.