Journal:

Article Title: Long Term Synaptic Depression That Is Associated with GluR1 Dephosphorylation but Not ?-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid (AMPA) Receptor Internalization *

doi: 10.1074/jbc.M803431200

Figure Lengend Snippet: Neither GluR1 nor GluR2/3 total surface expression is changed 45 min following cLTD treatment, as determined by cross-linking assay. A, representative blots of X-linked (25.0 μg of total protein loaded) and un-X-linked (5.0 μg of total protein loaded) samples for control and cLTD conditions probed with antibodies specific for GluR1 (left group) and GluR2/3 (right group). B, representative blots of un-X-linked and X-linked (5.0μg of total protein loaded for both) samples probed with antibodies specific for PP2Aα (top bands) and synapsin (bottom bands). C and D, graphs of cumulative data comparing X-linked control and X-linked cLTD samples probed with antibodies specific for GluR1 (C) and GluR2/3 (D)(n = 8). E and F, data from C and D normalized to the transferrin receptor. G and H, graphs of cumulative data comparing the percent surface expression (see “Experimental Procedures” for explanation of calculation) between control and cLTD conditions for GluR1 (G) and GluR2/3 (H)(n = 8). I and J, graphs of cumulative data comparing un-X-linked and X-linked samples probed with antibodies specific for PP2Aα (I) and synapsin (J)(n = 8). No significant differences were observed for any of the comparisons as determined by Student's paired t test. Values represent the mean ± S.E.

Article Snippet: Primary antibodies were as follows: GluR1 and GluR2/3 (Chemicon) used at 1:3000 in BSA, phospho-Ser-845 and phospho-Ser-831 (PhosphoSolutions) used at 1:1000 in BSA, PP2Aα (BD Pharmingen) used at 1:5000 in milk, synapsin, NR2B, and NR1 (PhosphoSolutions) used at 1:2000 in BSA, the α subunit of calcium/calmodulin-dependent protein kinase II (αCaMKII) (BD Pharmingen) used at 1:3000 in BSA, phospho-Thr-286 (PhosphoSolutions) used at 1:3000 in milk, PSD-95 (Affinity Bioreagents) used at 1:5000 in BSA, and transferrin receptor (Zymed Laboratories Inc.) used at 1:1000 in milk.

Techniques: Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cells Restore Endothelial Integrity and Alleviate Emotional Impairments in a Diabetic Mouse Model via Inhibition of MMP-9 Activity

doi: 10.3390/ijms26073355

Figure Lengend Snippet: Figure 2. DM mice exhibit synaptic deficits and increased neuroinflammation in the HIP. (A) Repre- sentative WB images showing the expression levels of the presynaptic protein synapsin I (SYN1) and the postsynaptic protein postsynaptic density protein 95 (PSD95) in the HIP of CTL and DM mice. β-actin was used as an internal control. (B) Semi-quantitative analysis of PSD95 and SYN1 expression levels from immunoblot experiments. n = 6. (C) Representative images of Iba1 immunostaining in the HIP of CTL and DM mice. The area within the dashed box is magnified and displayed in the insets. Scale bars: 100 µm (main images) and 20 µm (insets). (D) Quantitative analysis of Iba1-positive microglia in the HIP of CTL and DM mice. n = 5. (E) qPCR analysis of iNOS and Arg1 mRNA levels in the HIP of CTL and DM mice. n = 6. (F) qPCR analysis of IL-6, IL-1β, and TNF-α mRNA levels in the HIP of CTL and DM mice. n = 6. (G) ELISA measuring IL-6, IL-1β, and TNF-α levels in the serum of CTL and DM mice. n = 3. Data are presented as mean ± SEM. Statistical significance was determined using a one-tailed unpaired t-test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The membranes were blocked and incubated overnight with the following antibodies: PSD95 (SYSN, Toronto, ON, Canada, 124002, 1:1000); SYN1 (SYSN, 106011, 1:1000); Cldn5 (Invitrogen, 35-2500, 1:1000); Ocln (Invitrogen, 71-1500, 1:1000); and MMP9 (Boster, Shanghai, China; PB9669, 1:1000).

Techniques: Expressing, Control, Western Blot, Immunostaining, Enzyme-linked Immunosorbent Assay, One-tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Mesenchymal Stem Cells Restore Endothelial Integrity and Alleviate Emotional Impairments in a Diabetic Mouse Model via Inhibition of MMP-9 Activity

doi: 10.3390/ijms26073355

Figure Lengend Snippet: Figure 6. MSC treatment attenuated DM-induced neuroinflammation and synaptic deficits. (A) Representative immunofluorescence images of Iba1 staining in the HIP of mice from the CTL, MSC, DM, and DM+MSC groups. The dashed box indicates the magnified region shown in the insets. Scale bars: 100 µm (main images) and 20 µm (insets). (B) Quantitative analysis of Iba1+ microglia in the HIP. n = 4. (C) qPCR analysis of iNOS and Arg1 mRNA levels in the HIP of mice from the CTL, MSC, DM, and DM+MSC groups. n = 9. (D) qPCR analysis of mRNA levels of IL-6, IL-1β, and TNF-α in the HIP of mice from the CTL, MSC, DM, and DM+MSC groups. n = 9. (E) ELISA results showing serum levels of IL-6, IL-1β, and TNF-α in CTL, MSC, DM, and DM+MSC mice. n = 4. (F) Representative WB images of PSD95 and SYN1 expression in the HIP. β-actin was used as a loading control. (G) Semi-quantitative analysis of PSD95 and SYN1 protein levels. n = 6. Data are presented as mean ± SEM. Two-way ANOVA followed by Tukey’s post hoc test. ** p < 0.01, *** p < 0.001.

Article Snippet: The membranes were blocked and incubated overnight with the following antibodies: PSD95 (SYSN, Toronto, ON, Canada, 124002, 1:1000); SYN1 (SYSN, 106011, 1:1000); Cldn5 (Invitrogen, 35-2500, 1:1000); Ocln (Invitrogen, 71-1500, 1:1000); and MMP9 (Boster, Shanghai, China; PB9669, 1:1000).

Techniques: Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Control