Ysed onPage 2 of(page number not for citation purposes)Cell Communication and Signaling 2005, 3:http://www.biosignaling.com/content/3/1/Figure PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/25957400 1 TPO is elevated in AMM patients TPO is elevated in AMM patients. TPO level in the Procyanidin B1 cost undiluted Platelet Poor Plasma samples of 10 controls and 20 AMM patients was tested by ELISA. While TPO level was in the range of 0?1 pg/ml in controls (Mean 22.8 pg/ml), it ranged between 0?84 pg/ml in AMM patients (Mean 229 pg/ ml). This elevation of TPO level in patients was significant (P < 0.001) compared to controls.Figure 3 Mpl protein is reduced in AMM patients Mpl protein is reduced in AMM patients. Platelet lysates from 12 controls and 12 AMM patients in 1X Laemelli buffer were analysed by 6 SDS PAGE. Mpl antibody raised against the full-length protein was used for western blot analysis. The blots were re-probed with Actin Antibody for normalization. This representative autoradiogram shows the results for 4 controls and 7 AMM patients.SDS PAGE. Mpl antibody that was raised against the full length Mpl protein was used for western blot analysis. This antibody recognizes both glycosylated and non-glycosylated forms of Mpl. The blots were re-probed with Actin antibody for normalization. The western blots on chemiluminescence detection showed an overall reduction in the expression of Mpl in the platelets of patients with AMM and there was no co-relation (r2 = 0.0038) between TPO level and Mpl levels (Fig. 2). While there was high expression of glycosylated form of Mpl in the controls, the expression of Mpl in the platelets of patients with AMM varied from none to having reduced amounts of both glycosylated and faster moving non-glycosylated forms. A representative blot with results from 4 controls and 7 patients is shown in Fig. 3. The developed blots were analysed using the Image J program to quantitate the intensity of the bands and the values for Mpl expression after normalization against Actin expression are represented in Fig. 4. Statistical analysis of the values by t-test revealed the reduced Mpl expression to be significant (P < 0.05).Mpl RNA level is similar in controls and AMM patients Since Mpl protein level was reduced in AMM patients, to know if it was due to reduced transcription of the Mpl gene, we compared the levels of Mpl RNA in AMM patients and controls by Real-Time RT-PCR. Total RNA from the platelets of 12 controls and 12 AMM patients were used as templates for PCR with gene specific primers. The values were normalized against GAPDH expression and are represented in Fig. 5. On statistical analysis (ttest), the difference in Mpl expression was found not to beFigure 2 TPO level is not co-related to platelet Mpl protein level TPO level is not co-related to platelet Mpl protein level. Plasma TPO levels and platelet Mpl protein levels from 12 AMM patients were compared for co-relation. Statistical analysis revealed absence of any co-relation between the levels of these two proteins (r2 = 0.0038).Page 3 of(page number not for citation purposes)Cell Communication and Signaling 2005, 3:http://www.biosignaling.com/content/3/1/Figure 4 Densitometric scanning results of Mpl expression in platelets Densitometric scanning results of Mpl expression in platelets. The intensity of the bands on the Mpl western blot autoradiograph was quantitated using Image J program. The values represent Mpl expression after normalization with Actin expression assayed for 12 controls and 12 AMM patients. The reduct.