Journal: International journal of molecular sciences

Article Title: Phosphodiesterase 5a Signalling in Skeletal Muscle Pathophysiology.

doi: 10.3390/ijms24010703

Figure Lengend Snippet: Figure 4. Modulation of Pde5a1 by MYOD and RUNX1. (a) Scheme of Pde5A promoter constructs transfected in cells. C2C12 cells were transiently co-transfected with a plasmid expressing MyoD (b) or Runx1 (c) and the different Pde5a1 promoter constructs. Results are the means of at least three different experiments performed in triplicate and expressed as the fold change activity relative to that observed in cells not overexpressing MyoD or Runx1. (d) C2C12 cells were transiently transfected with a plasmid expressing MyoD or Runx1 and the levels of Pde5a1 RNA were measured by RT- qPCR. Relative expression of Pde5a1 in cells overexpressing MYOD (dotted column) or RUNX1 (grey column) are expressed as the fold change compared to untreated cells (empty columns) All values represent the mean ± SD of n = 3 different RNA preparations. * Indicates statistical significance by the t-test (p < 0.05).

Article Snippet: 2023, 24, 703 11 of 14 pression vectors coding for MyoD and Runx1 were purchased from Addgene (Watertown, MA, USA).

Techniques: Construct, Transfection, Plasmid Preparation, Expressing, Activity Assay, Quantitative RT-PCR

Journal: International journal of molecular sciences

Article Title: Phosphodiesterase 5a Signalling in Skeletal Muscle Pathophysiology.

doi: 10.3390/ijms24010703

Figure Lengend Snippet: Figure 5. MyoD and Runx1 mRNA levels in skeletal muscle under pathophysiological conditions. The mRNA levels of MyoD and Runx1 were measured by RT-qPCR. mRNA levels in the samples (dotted or grey columns) are expressed as the fold change compared with the samples from muscle from wild-type mice (empty columns). All values represent the mean ± SD of n = 3 different RNA preparations. * Indicates statistical significance of pathological samples compared with control samples (t-test: p < 0.05).

Article Snippet: 2023, 24, 703 11 of 14 pression vectors coding for MyoD and Runx1 were purchased from Addgene (Watertown, MA, USA).

Techniques: Quantitative RT-PCR, Control

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Engineering an inducible leukemia-associated fusion protein enables large-scale ex vivo production of functional human phagocytes.

doi: 10.1073/pnas.2312499121

Figure Lengend Snippet: Fig. 1. Establishment of a protein switch for human hematopoietic stem and progenitor cell expansion using the MLL-ENL (ME) fusion gene and the Shield1 destabilization domain (DD). (A) Illustration of the procedure by which functional monocytes were derived from transduced and expanded human bone marrow CD34+ cells. (B) Gene ontology (GO) analysis of ME target genes (36) categorized by function (33–35). (C) A depiction of the retroviral vector containing ME fused to a DD destabilization domain (DD-ME). (D) Transduced CD34+ cells cultured in the presence of IL-3, IL-6, SCF, TPO, Flt3-L, and GM-CSF with or without Shield1 and analyzed for eGFP expression. (E) FSC/SSC profiles of DD-ME-expressing cells in the presence of Shield1 at day 44 posttransduction. (F) DD-ME signal intensity in the presence of Shield1 (+S) and after withdrawal of Shield1 (for 7 d) via intracellular staining. The data are presented as the mean plus the SD of three biological replicates. (G) Western blot analysis of HOXA9 expression using whole cell lysates of DD-ME-expressing cells before and after Shield1 removal. (H) Outgrowth of transduced, eGFP-expressing cells was determined via flow cytometry. #, terminally differentiated; FSC, forward scatter; SSC, side scatter.

Article Snippet: The sequences of HOXB8 (synthesized by Geneart) and HOXA9 (Addgene #97041) were cloned into the same retroviral backbone.

Techniques: Functional Assay, Derivative Assay, Retroviral, Plasmid Preparation, Cell Culture, Expressing, Staining, Western Blot, Flow Cytometry