Journal: Molecular Therapy

Article Title: siRNA for REST ameliorates symptoms in ALS mice and serum REST predicts disease prognosis and survival in ALS patients

doi: 10.1016/j.ymthe.2025.10.039

Figure Lengend Snippet: Serum levels and prognostic value of REST protein in ALS patients (A) ELISA assay results showing REST protein levels in the serum of ALS patients ( n = 56), compared with healthy controls (Ctrl) ( n = 23). ∗∗ p < 0.01 vs. control by Student’s t test. (B) Kaplan-Meier analysis of tracheostomy-free survival in ALS patients stratified according to the median serum REST protein levels. Patients with REST levels above the median, represented in red, exhibit a significantly worse prognosis than patients with levels below the median, shown in light green. (C and D) Univariate and multivariate Cox proportional hazard models assessing the prognostic value of increased serum REST levels, adjusted for bulbar onset type, sex, progression rate, and age at serum sampling. Whiskers represent the 95% confidence interval (CI). ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Serum REST level was quantified by a human-specific ELISA kit specific for REST (Cusabio, American Research Products, Waltham, MA).

Techniques: Enzyme-linked Immunosorbent Assay, Control, Sampling

Journal: Antioxidants (Basel, Switzerland)

Article Title: Hydrogen Peroxide-Induced Re-Expression of Repressor Element 1-Silencing Transcription Factor Contributes to Cardiac Vagal Dysfunction in Type 2 Diabetes Mellitus.

doi: 10.3390/antiox14050588

Figure Lengend Snippet: Figure 3. Effect of REST shRNA transfection on N-type Ca2+ channel (Cav2.2-α) protein expression in AVG neurons from T2DM rats. (A) Raw and quantitative data representing the REST protein levels in AVG neurons from all groups of rats, analyzed by reverse-phase protein microarray. (B) Raw and quantitative data representing Cav2.2-α protein levels in the AVG neurons from all groups of rats. Black dots represent each individual data point. N = 12 measurements from 6 rats/group. Data are means ± SEM. One-way ANOVA with post-hoc Bonferroni test was used to assess statistical significance. * p < 0.05 vs. sham; # p < 0.05 vs. T2DM.

Article Snippet: Under the microscope, 2 μL of saline, adenoviral catalase gene (Ad.CAT, 1 × 1010 pfu/mL, University of Iowa, Iowa City, IA, USA), adenoviral vector control (Ad.Empty, 1 × 1010 pfu/mL, University of Iowa, Iowa City, IA, USA), rat lentiviral REST shRNA (Lenti.REST shRNA, 29mer target-specific shRNA designed against multiple slice variants based on NM_031788 and NM_031788.1, 1 × 107 TU/mL, CAT#: TL711581V, OriGene Technologies, Inc., Rockville, MD, USA), or lentiviral scrambled shRNA (Lenti.scrambled shRNA, lenti particles carrying a 29mer scrambled shRNA sequence cassette, 1 × 107 TU/mL, CAT#: TR30021V, OriGene Technologies, Inc., Rockville, MD, USA) was microinjected into the AVG by a glass micropipette connected to a WPI Nanoliter 2000 microinjector (World Precision Instruments, Sarasota, FL, USA).

Techniques: shRNA, Transfection, Expressing, Microarray

Journal: Antioxidants (Basel, Switzerland)

Article Title: Hydrogen Peroxide-Induced Re-Expression of Repressor Element 1-Silencing Transcription Factor Contributes to Cardiac Vagal Dysfunction in Type 2 Diabetes Mellitus.

doi: 10.3390/antiox14050588

Figure Lengend Snippet: Figure 4. Effect of REST shRNA transfection on N-type Ca2+ currents in AVG neurons from T2DM rats. (A) Raw data for Ca2+ currents from sham, T2DM, and T2DM + lenti.REST shRNA rats. (B) Quantitative data for total Ca2+ currents and other types of Ca2+ currents (cells treated with ω-conotoxin GVIA, a specific N-type Ca2+ channel blocker) recorded under the test pulse at 0 mV in AVG neurons from all groups of rats. (C) Quantitative data for N-type Ca2+ currents analyzed from the original recording in AVG neurons from all groups of rats. Subtracting Ca2+ currents under treatment of ω-conotoxin GVIA from total Ca2+ currents in the original recording was used to obtain N-type Ca2+ currents. Black dots represent each individual data point. N = 6 neurons from 4 rats/group; data are means ± SEM. One-way ANOVA with post-hoc Bonferroni test was used to assess statistical significance. * p < 0.05 vs. sham; # p < 0.05 vs. T2DM.

Article Snippet: Under the microscope, 2 μL of saline, adenoviral catalase gene (Ad.CAT, 1 × 1010 pfu/mL, University of Iowa, Iowa City, IA, USA), adenoviral vector control (Ad.Empty, 1 × 1010 pfu/mL, University of Iowa, Iowa City, IA, USA), rat lentiviral REST shRNA (Lenti.REST shRNA, 29mer target-specific shRNA designed against multiple slice variants based on NM_031788 and NM_031788.1, 1 × 107 TU/mL, CAT#: TL711581V, OriGene Technologies, Inc., Rockville, MD, USA), or lentiviral scrambled shRNA (Lenti.scrambled shRNA, lenti particles carrying a 29mer scrambled shRNA sequence cassette, 1 × 107 TU/mL, CAT#: TR30021V, OriGene Technologies, Inc., Rockville, MD, USA) was microinjected into the AVG by a glass micropipette connected to a WPI Nanoliter 2000 microinjector (World Precision Instruments, Sarasota, FL, USA).

Techniques: shRNA, Transfection

Journal: Antioxidants (Basel, Switzerland)

Article Title: Hydrogen Peroxide-Induced Re-Expression of Repressor Element 1-Silencing Transcription Factor Contributes to Cardiac Vagal Dysfunction in Type 2 Diabetes Mellitus.

doi: 10.3390/antiox14050588

Figure Lengend Snippet: Figure 5. Effect of REST shRNA transfection on neuronal excitability in AVG neurons from T2DM rats. Raw (A) and quantitative data (B) for action potentials during a 1 s current clamp with a current injection of 100 pA in AVG neurons from all groups of rats. Black dots represent each individual data point. N = 6 neurons from 4 rats/group; data are means ± SEM. One-way ANOVA with post-hoc Bonferroni test was used to assess statistical significance. * p < 0.05 vs. sham; # p < 0.05 vs. T2DM.

Article Snippet: Under the microscope, 2 μL of saline, adenoviral catalase gene (Ad.CAT, 1 × 1010 pfu/mL, University of Iowa, Iowa City, IA, USA), adenoviral vector control (Ad.Empty, 1 × 1010 pfu/mL, University of Iowa, Iowa City, IA, USA), rat lentiviral REST shRNA (Lenti.REST shRNA, 29mer target-specific shRNA designed against multiple slice variants based on NM_031788 and NM_031788.1, 1 × 107 TU/mL, CAT#: TL711581V, OriGene Technologies, Inc., Rockville, MD, USA), or lentiviral scrambled shRNA (Lenti.scrambled shRNA, lenti particles carrying a 29mer scrambled shRNA sequence cassette, 1 × 107 TU/mL, CAT#: TR30021V, OriGene Technologies, Inc., Rockville, MD, USA) was microinjected into the AVG by a glass micropipette connected to a WPI Nanoliter 2000 microinjector (World Precision Instruments, Sarasota, FL, USA).

Techniques: shRNA, Transfection, Injection

Journal: Antioxidants (Basel, Switzerland)

Article Title: Hydrogen Peroxide-Induced Re-Expression of Repressor Element 1-Silencing Transcription Factor Contributes to Cardiac Vagal Dysfunction in Type 2 Diabetes Mellitus.

doi: 10.3390/antiox14050588

Figure Lengend Snippet: Figure 6. Effect of REST shRNA on vagal control of the ventricular function in T2DM. Left vagal efferent nerve stimulation (VNS)-induced changes of the left ventricular systolic pressure (LVSP, panel A) and the maximum rate of increase of left ventricular pressure (LV dP/dtmax, panel B) represented the vagal control of the ventricular function. N = 6 rats/group; data are means ± SEM. Two-way repeated measures ANOVA with post-hoc Bonferroni was used to test statistical significance. * p < 0.05 vs. sham; # p < 0.05 vs. T2DM.

Article Snippet: Under the microscope, 2 μL of saline, adenoviral catalase gene (Ad.CAT, 1 × 1010 pfu/mL, University of Iowa, Iowa City, IA, USA), adenoviral vector control (Ad.Empty, 1 × 1010 pfu/mL, University of Iowa, Iowa City, IA, USA), rat lentiviral REST shRNA (Lenti.REST shRNA, 29mer target-specific shRNA designed against multiple slice variants based on NM_031788 and NM_031788.1, 1 × 107 TU/mL, CAT#: TL711581V, OriGene Technologies, Inc., Rockville, MD, USA), or lentiviral scrambled shRNA (Lenti.scrambled shRNA, lenti particles carrying a 29mer scrambled shRNA sequence cassette, 1 × 107 TU/mL, CAT#: TR30021V, OriGene Technologies, Inc., Rockville, MD, USA) was microinjected into the AVG by a glass micropipette connected to a WPI Nanoliter 2000 microinjector (World Precision Instruments, Sarasota, FL, USA).

Techniques: shRNA, Control

Journal: Journal of Neuroscience

Article Title: Long Noncoding RNA FosDT Promotes Ischemic Brain Injury by Interacting with REST-Associated Chromatin-Modifying Proteins

doi: 10.1523/jneurosci.2943-15.2015

Figure Lengend Snippet: Figure2. TransientMCAOinducedFosDTexpressionintheipsilateralcortexat3,6,and12hofreperfusioncomparedwithshamanalyzedwithLncRNAmicroarrays(A).IncreasedFosDTandFos expressionwasobservedat12hofreperfusionaftertransientMCAOwithreal-timePCR(B).LevelsofacontrolLncRNAwerenotalteredatthistimepoint(B).ExpressionofRESTanditscorepressor CMPs coREST and MeCP2 measured by real-time PCR was observed to be increased at 12 h reperfusion following transient MCAO (C). Protein levels of REST and coREST were also observed to be increased at 12 h reperfusion following transient MCAO (D, E). RIP showed increased binding of coREST and Sin3a to FosDT at 6 h of reperfusion following transient MCAO (F). Values are mean SD (n 3 or 4/group).*p 0.05, compared with sham by Student’s t test.

Article Snippet: In brief, tissue from peri-infarct area was homogenized in buffer containing protease and phosphatase inhibitors, centrifuged at 14,000 g, and the supernatant (40 g) was electrophoresed, blotted onto nitrocellulose membranes, and probed with antibodies against REST (1:500; ARP32478P050, Aviva System Biology), coREST (1:5000; ab183711, Abcam), Sin3a (1:1000; ab129087, Abcam), and -actin (1:000; 3700, Cell Signaling Technology) followed by HRP-conjugated anti-rabbit or anti-mouse IgG (Cell Signaling Technology).

Techniques: Real-time Polymerase Chain Reaction, Binding Assay