Journal: Haematologica

Article Title: Impact of Sox11 over-expression in Ba/F3 cells

doi: 10.3324/haematol.2018.197467

Figure Lengend Snippet: Phenotypical, proliferative and transcriptional changes in Ba/F3 cells upon 72 h of induced Sox11 expression. (A) Western Blot of SOX11 protein expression in Sox11-ON (doxycycline supplemented medium) and Sox11-OFF (control medium) cells at 72 h, detected with the rabbit polyclonal anti-SOX11 antibody HPA000536, Sigma-Aldrich. B) Bright field microscopy images of cell aggregates following 72 h of continuous Sox11 expression (10x), imaged by Nikon Ti-E microscope. C) Sox11-ON cells incorporates less 3H-Thymidine at 72 h of Sox11 induction following a 4 h pulse, as compared to Sox11-OFF cells, measured in counts per minute, error bars represent the standard deviation (P=0.0086, n=3). D) Mean relative absorption measured by XTT (n=3). Metabolic activity is significantly reduced (P=0.0124) in Sox11-ON cells as compared to Sox11-OFF cells. E) Volcano plot representation of transcript level differences by Affymetrix MTA-1 mouse arrays (Microarray data has been made available through the GEO database with accession number GSE108419). Names are shown for the genes with the largest transcript level fold change (log2FC ≥1.3 or ≤−1.3). Blue and red: genes with significantly altered transcript levels in Sox11-ON cells and a fold change below -2 (blue) (FDR q-value ≤0.05 and log2FC ≥−1) or above 2 (red) (FDR q-value ≤0.05 and log2FC ≥1); gray: genes significantly changed at the transcript level (FDR q-value ≤0.05); black: genes with non-significant transcript level changes. F) Expression levels after Sox11 induction in genes specifically expressed at different stages of B-cell development. Only the pro-B restricted genes Id1 and Tal1 had significantly altered transcript levels in Sox11-ON cells (FDR q-value: 0.006 and 0.016, respectively). None of the other investigated pro-B and pre-B cell associated genes were altered at the transcript level. Genes associated with later B-cell developmental stages are shown for comparison. Transcript levels are presented as a gene-wise standardized expression (Z-score). FC: fold change.

Article Snippet: E) Volcano plot representation of transcript level differences by Affymetrix MTA-1 mouse arrays (Microarray data has been made available through the GEO database with accession number GSE108419).

Techniques: Expressing, Western Blot, Microscopy, Standard Deviation, Activity Assay, Microarray

Journal: Frontiers in Cell and Developmental Biology

Article Title: Biallelic Mutations in ACACA Cause a Disruption in Lipid Homeostasis That Is Associated With Global Developmental Delay, Microcephaly, and Dysmorphic Facial Features

doi: 10.3389/fcell.2021.618492

Figure Lengend Snippet: Summary of the genetic characteristics and SDS-PAGE of ACC1 protein content and ACC1 enzyme activity. (A) The pedigree of the affected individual. The proband was represented as black solid circle and pointed out by an arrow. The square indicates male, and the circle female. (B) Sanger sequencing of the mutations. (C) The amino acid conservation among different species. (D) The functional domain of ACC1, and the mutation sites were marked. (E) Relative ACC1 protein content of lymphocytes. (F) The ACC1 enzyme activity among the proband- and age-matched control-derived lymphocytes. One-way ANOVA applied in panels (E,F) . ** p < 0.01.

Article Snippet: According to the Human Protein Atlas, ACC1 expression level in fibroblasts is almost 66.7% compared to HUVEC, however, in our data the expression level in fibroblasts and lymphocytes was even higher than HUVEC ( ).

Techniques: SDS Page, Activity Assay, Sequencing, Functional Assay, Mutagenesis, Control, Derivative Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Biallelic Mutations in ACACA Cause a Disruption in Lipid Homeostasis That Is Associated With Global Developmental Delay, Microcephaly, and Dysmorphic Facial Features

doi: 10.3389/fcell.2021.618492

Figure Lengend Snippet: ACC1-knockdown (KD) fibroblasts and the cell motility capacity measurement. (A) The ACC1 protein content of ACC1-KD fibroblast cell lines, and (B) the ACC1 enzyme activity. The wound healing assay was carried out at different time points (0, 12, and 24 h) (C) without or (E) with palmitate (scale bar = 30 μm), and (D,F) show quantitative analysis, respectively. The Trans-well migration capacity at 100× magnification (upper) and 200× magnification (lower) (G) with or (I) without palmitate, respectively. (H,J) The quantitative analyses, respectively. All data were from three independent replicates and analyzed by mean ± SEM. Independent t -test in panels (A,B,D,F,H,J) . * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: According to the Human Protein Atlas, ACC1 expression level in fibroblasts is almost 66.7% compared to HUVEC, however, in our data the expression level in fibroblasts and lymphocytes was even higher than HUVEC ( ).

Techniques: Knockdown, Activity Assay, Wound Healing Assay, Migration