Figure 3
Expression levels of SDIM1 protein in human and mouse tissues. A. Anti-SDIM1 antibody was tested on recombinant GST-SDIM1 fusion protein (right lane). Left lane-Coomassie Blue stained GST-SDIM1. B. Anti-SDIM1 antibody detected SDIM1 in proteins extracted from NT2 neurons. C. SDIM1 protein from mouse brain (lane 1), muscle (lane 2) and HEK-293 cells over-expressing SDIM1 (lane 3) was detected with anti-SDIM1 (left panel) or anti-ubiquitin (right panel) antibody. Lane 4-purified human ubiquitin. D. GST-SDIM1 protein was mixed with proteins from HEK-293 cells transfected with pEGFP-N1 or pSDIM1-EGFP and immunoprecipitated with anti-GST antibody. The presence of monomeric and dimeric GST-SDIM1, GST-SDIM/SDIM1-EGFP or SDIM1-EGFP was detected by anti-GST (left panel) or anti-EGFP (middle panel) antibody. The second blot was stripped and probed with anti-SDIM1 anti-body (right panel). Lane1-total proteins from pEGFP-N1 transfected cells, lane 2-total proteins from pSDIM1-EGFP transfected cells, lane 3-mixture of GST-SDIM1 and protein from pEGFP-N1 transfected cells immunoprecipitated with anti-GST antibody, lane 4-mixture of GST-SDIM1 and protein from pSDIM1-EGFP transfected cells immunoprecipitated with anti-GST antibody, lane 5-mock immunoprecipitation without anti-GST antibody. E. The expression levels of SDIM1 protein in human tissues were detected by Western analysis with anti-SDIM1 antibody. A representative blot from each lot was tested by the vendor with anti-GAPDH antibody. Lane B-Brain, lane K-Kidney, lane L-Lung, lane S-Spleen, lane I-Small Intestine, lane U-Uterus, lane C-Cervix, lane P-Placenta. F. SDIM1 protein in mouse tissues was detected by Western analysis with anti-SDIM1 antibody. The striped membrane was re-blotted with anti-β-actin and anti-GAPDH antibodies. Lane B-Brain, lane I-Intestine, lane C-Cervix, lane Li-Liver, lane M-Muscle, lane H-Heart, lane L-Lung, lane K-Kidney, lane S-Spleen. G. SDIM1 protein in control and AD human brains were detected by Western analysis with anti-SDIM1 antibody.