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Results: 4

1.
Figure 2

Figure 2. From: Influence of microRNA deregulation on chaperone-mediated autophagy and α-synuclein pathology in Parkinson's disease.

Luciferase reporter assays to analyze the influence of miRNAs on the 3′UTR of lamp-2a and hsc70. (a) Schematic representation of the Renilla luciferase reporter constructs in psiCHECK2.2 for lamp-2a and hsc70 3′UTR. (b) The influence of increasing concentrations of hsa-miR-106a* and hsa-miR-224 on Renilla luciferase activity when cotransfected with luciferase-3′UTR hsc70 or luciferase-3′UTR lamp-2a constructs. (c) The effect of cotransfection of the different miRNAs (10 nM) with either luciferase-3′UTR hsc70 or luciferase-3′UTR lamp-2a reporter constructs upon Renilla luciferase activity 48 h after transfection. Data normalized to cells in the absence of miRNA (C). Values are mean±S.E.M. (n=6), statistical analyses compared with the respective control group, *P<0.05, **P<0.01, ***P<0.001

L Alvarez-Erviti, et al. Cell Death Dis. 2013 March;4(3):e545.
2.
Figure 3

Figure 3. From: Influence of microRNA deregulation on chaperone-mediated autophagy and α-synuclein pathology in Parkinson's disease.

Impact of increased miRNA levels upon LAMP-2A, hsc70 and α-synuclein protein levels. (a and b) Western blot analyses of normal SHSY5Y cells 72 h after transfection with the respective miRNAs (10 nM) and untreated cells (C) for (a) LAMP-2A, hsc70 and actin (b) and their quantitation. (c and d) Western blot analyses of α-synuclein overexpressing cells 9 days after treatment with miRNAs (10 nM) for (c) α-synuclein, LAMP-2A, hsc70 and actin. (d) Quantitation of α-synuclein, LAMP-2A and hsc70 protein levels. Values are relative to actin and normalized to untreated (control) cells. Data expressed as mean±S.E.M. (n=3), statistical analyses compared with the respective control group, *P<0.05

L Alvarez-Erviti, et al. Cell Death Dis. 2013 March;4(3):e545.
3.
Figure 4

Figure 4. From: Influence of microRNA deregulation on chaperone-mediated autophagy and α-synuclein pathology in Parkinson's disease.

Analysis of PD brain samples and the dose-dependent impact of miRNA-373* upon LAMP-2A. Relative change in miRNAs normalized to actin mRNA levels and compared with control in (a) SNc from PD patients and (b) the amygdala. (c) mRNA levels for lamp-2a, hsc70 and α-synuclein relative to actin mRNA and compared with control values in PD amygdala and SNc. Data expressed as mean±S.E.M., controls (n=5) and PD (n=6). Statistical analyses compared with control group, *P<0.05. (d) The influence of transfecting increasing concentrations of miRNA hsa-miR-373* into normal SH-SY5Y cells on LAMP-2A protein and mRNA levels 72 h after transfection. The upper panel depicts the western blot of LAMP-2A levels relative to actin with increasing miR-373* levels, and the lower panel depicts the relationship between LAMP-2A protein and lamp-2a mRNA levels relative to actin and normalized to untreated cells

L Alvarez-Erviti, et al. Cell Death Dis. 2013 March;4(3):e545.
4.
Figure 1

Figure 1. From: Influence of microRNA deregulation on chaperone-mediated autophagy and α-synuclein pathology in Parkinson's disease.

Influence of pathological features associated with PD on LAMP-2A, hsc70 and α-synuclein levels. (ac) Normal SH-SY5Y cells after 48 h of treatment with different concentrations of epoxomycin (E), paraquat (P), rotenone (R) or untreated (C). (a) Western blot analyses of hsc70, LAMP-2A and actin. (b) Quantitation of the western blot data for LAMP-2A (open bars) and hsc70 (closed bars) protein levels relative to actin and normalized to untreated cells. (c) qPCR analysis of hsc70 and LAMP-2A mRNA relative to actin mRNA and normalized to untreated cells. (df) SH-SY5Y cells overexpressing WT α-synuclein after 48 h of exposure to epoxomycin (E, 5 nM), paraquat (P, 100 μM), rotenone (R, 200 nM) or under normal conditions (C). (d) Western blot analyses and (e) quantitation of the western blot data for LAMP-2A (open bars) and hsc70 (closed bars) protein levels relative to actin and normalized to untreated (control) cells. (f) Quantitation of the western blot data for α-synuclein protein relative to actin and mRNA levels relative to actin mRNA; all data normalized to untreated (control) cells. Data expressed as mean±S.E.M. (n=3), and statistical analyses compared with the respective control group, *P<0.05

L Alvarez-Erviti, et al. Cell Death Dis. 2013 March;4(3):e545.

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