Journal: Journal of dermatology and skin science

Article Title: The Dawning of a New Enterprise: RNA Therapeutics for the Skin

doi: 10.29245/2767-5092/2023/1.1168

Figure Lengend Snippet: RNA-based therapies currently in clinical trials for skin conditions

Article Snippet: TD101 , Phase I , NCT00716014 , Completed , Pachyonychia Congenita , Transderm/International Pachyonychia , siRNA targeting one disease-causing mutation of keratin 6a (K6a), the N171K mutant , Pachyonychia congenita siRNA; Reveker; sdTD-K6a.513a.12; siRNA sdTD101; TD-K6a.513a.12; TD101.

Techniques: Clinical Proteomics, Expressing, Activity Assay, Mutagenesis, Knockdown, Functional Assay, Immunopeptidomics

Journal: Metabolic brain disease

Article Title: Effect of AmyTrap, an Amyloid-β Binding drug, on Aβ induced Mitochondrial dysfunction and Tau phosphorylation in cultured neuroblastoma cells

doi: 10.1007/s11011-019-00520-2

Figure Lengend Snippet: Each enzyme is represented by a representative immunoblot and graph of the band intensity (A) Pyruvate Dehydrogenase (43 kDa), (B) ATP Citrate Lyase (121 kDa), (C) Succinate Dehydrogenase (72 kDa), (D) Oxoglutarate Dehydrogenase (116 kDa). Treatment groups are Untreated, wild-type Aβ42 (4 μM), mutant AβY10A (4 μM) or a combination of Aβ42 (4 μM) and RI-peptide (10 μM). Band intensities are from 3 different experiments and values are represented as Mean ± SD. Significance was calculated by Student’s t-test against the untreated neuroblastoma group for all treatments and then between Aβ and AβY10A or Aβ and Aβ + RI-peptide. A * indicates p < 0.05, ** indicates p < 0.01 and *** indicates p < 0.001.

Article Snippet: Wild-type and mutant Aβ42 were purchased from Anaspec [CA] for these experiments.

Techniques: Western Blot, Mutagenesis

Journal: Metabolic brain disease

Article Title: Effect of AmyTrap, an Amyloid-β Binding drug, on Aβ induced Mitochondrial dysfunction and Tau phosphorylation in cultured neuroblastoma cells

doi: 10.1007/s11011-019-00520-2

Figure Lengend Snippet: Analysis of phosphorylated Tau was performed through ImageJ analysis. The images of the immunoblot of Tau is shown below the graph. Treatment groups are Untreated, wild-type Aβ42 (4 μM), mutant AβY10A (4 μM) or a combination of Aβ42 (4 μM) and RI-peptide (10 μM). Significance was calculated against Untreated Neuroblastoma group by Student’s t-test. Experiments were performed in triplicate. Values are represented as Mean ± SD. A * indicates p < 0.05, ** indicates p < 0.01.

Article Snippet: Wild-type and mutant Aβ42 were purchased from Anaspec [CA] for these experiments.

Techniques: Western Blot, Mutagenesis

Journal: Metabolic brain disease

Article Title: Effect of AmyTrap, an Amyloid-β Binding drug, on Aβ induced Mitochondrial dysfunction and Tau phosphorylation in cultured neuroblastoma cells

doi: 10.1007/s11011-019-00520-2

Figure Lengend Snippet: Analysis of the presence of intracellular RI-peptide was performed through ImageJ. Treatment groups are Untreated, RI-peptide (10 μM), or a combination of Aβ42 (4 μM) and RI-peptide (10 μM). A positive control consisting of a known concentration of RI-peptide (100 ng) was utilized to evaluate the intracellular concentration of peptide observed between treatment groups. The image of the immunoblot of RI-peptide is shown below the graph. Significance was calculated against Untreated Neuroblastoma group by Student’s t-test. Experiments were performed in triplicate. Values are represented as Mean ± SD. A ** indicates p < 0.01.

Article Snippet: Wild-type and mutant Aβ42 were purchased from Anaspec [CA] for these experiments.

Techniques: Positive Control, Concentration Assay, Western Blot

Journal: bioRxiv

Article Title: Hemagglutinin of Influenza A, but not of Influenza B and C viruses is acylated by ZDHHC2, 8, 15 and 20

doi: 10.1101/2019.12.15.877001

Figure Lengend Snippet: HeLa cells were transfected with siRNAs against all human ZDHHCs and 48 hours later transfected with a plasmid encoding H7 subtype HA from a variant of FPV. After 24 hours, cells were lysed and subjected to acyl-RAC and immune-blotting (IB). First with antiserum against the HA2 subunit (upper panel), then the membrane was again blotted with antibody against caveolin-1 as cellular control (lower panel). NH2OH: samples treated (+) or not treated (-) with hydroxylamine to cleave thioester-bound fatty acids. Input: western blot of 10% of the lysate to check expression levels of HA. 78 and 25 indicates the mobility of the molecular weight marker. The blots on the left were exposed for the same time as the blots in the middle and on the right.

Article Snippet: 48 hours later cells were transfected with a plasmid encoding HA of the FPV mutant using Fugene (Promega).

Techniques: Transfection, Plasmid Preparation, Variant Assay, Membrane, Control, Western Blot, Expressing, Molecular Weight, Marker

Journal: bioRxiv

Article Title: Hemagglutinin of Influenza A, but not of Influenza B and C viruses is acylated by ZDHHC2, 8, 15 and 20

doi: 10.1101/2019.12.15.877001

Figure Lengend Snippet: A: Phylogenetic tree of human ZDHHCs showing that ZDHHC2, 15 and 20 as well as 5 and 8 belong to the same group. B: [ 3 H]-palmitate labelling: HeLa cells were transfected with siRNAs against ZDHHC1, 2, 8, 15, with a scrambled siRNA (scra) or remain un-transfected (-) and 48h later with a plasmid encoding HA from H7 subtype from FPV. After 24 hours, cells were labeled for 2 hours with [ 3 H]-palmitate, lysed and subjected to immunoprecipitation with antiserum against the HA2 subunit (lower panel). An aliquot of the sample was subjected to western blotting with the same antibody (upper panel). The lower band in the upper panel is the heavy chain of the antibodies used for immuno-precipitation.

Article Snippet: 48 hours later cells were transfected with a plasmid encoding HA of the FPV mutant using Fugene (Promega).

Techniques: Transfection, Plasmid Preparation, Labeling, Immunoprecipitation, Western Blot

Journal: bioRxiv

Article Title: Hemagglutinin of Influenza A, but not of Influenza B and C viruses is acylated by ZDHHC2, 8, 15 and 20

doi: 10.1101/2019.12.15.877001

Figure Lengend Snippet: A549 human lung cells were transfected with plasmids encoding H7 subtype HA and the indicated ZDHHCs fused to a C-terminal HA-tag. 24 hours later cells were fixed, permeabilized and stained with anti-FPV antiserum, and anti-HA-tag antibodies followed by secondary antibody coupled to Alexa-Fluor 568 (red for ZDHHCs) and Alexa Fluor 488 (green for HA), respectively. Nuclei were stained with DAPI. Scale bar =50µm. Cells transfected with a plasmid expressing GST and stained with both antibodies was used as a negative control. Co-localization of HA with each ZDHHC from at least 40 cells was quantified with the Pearson’s correlation coefficient method using the JACoP plugin of the ImageJ software. 60% of HA pixels overlapped with ZDHHC8, 72% with ZDHHC2, 74% with ZDHHC20 and 75% with ZDHHC15.

Article Snippet: 48 hours later cells were transfected with a plasmid encoding HA of the FPV mutant using Fugene (Promega).

Techniques: Transfection, Staining, Plasmid Preparation, Expressing, Negative Control, Software