Journal: Scientific Reports

Article Title: CEBPA-dependent HK3 and KLF5 expression in primary AML and during AML differentiation

doi: 10.1038/srep04261

Figure Lengend Snippet: (a) NB4 or HT93 APL cell lines were stably transduced with pLKO.1 lentiviral vectors expressing non-targeting or two independent CEBPA -targeting shRNAs. APL control and CEBPA knockdown cells were differentiated with 1 μM ATRA for 4 days. Knockdown efficiency in NB4 (top panels) or HT93 (bottom panels) APL cells was validated by qPCR. CEBPA mRNA expression was normalized to the housekeeping gene HMBS and is shown as n-fold changes compared to untreated SHC002 control cells. HK3 (b) and KLF5 (c) mRNA expression in NB4 or HT93 APL cells was determined by qPCR and analyzed as in 2a. Impairment in granulocytic differentiation of CEBPA knockdown cells was shown by a reduction of the neutrophil marker granulocyte colony-stimulating factor receptor (GCSFR or CSF3R) (d) Data represent the mean ± s.d. of at least three independent experiments. (e), (f) CEBPA Knockdown efficiency at the protein level in NB4 and HT93 APL cells was confirmed by western blotting. GAPDH is shown as a loading control. MWU: *p < 0.05, **p < 0.01 and ***p < 0.001.

Article Snippet: Data represent log 2 expression levels and the measured cycle threshold (Ct) values were normalized to the expression levels of the housekeeping genes HMBS and ABL1 .For quantification of KLF5, CEBPA, CEBPE and G-CSFR mRNA in human cells we used Taqman® Gene Expression Assays Hs00156145_m1, Hs00269972_s1, Hs00357657_m1, and Hs00167918_m1, (Applied Biosystems), respectively.

Techniques: Stable Transfection, Transduction, Expressing, Control, Knockdown, Marker, Western Blot

Journal: Blood Advances

Article Title: JAK/STAT blockade reverses the malignant phenotype of Hodgkin and Reed-Sternberg cells

doi: 10.1182/bloodadvances.2021006336

Figure Lengend Snippet: Downregulation of CSF3R expression. (A) CSF3R GE changes over time (log 2 -fold change) in cHL-derived cell lines compared with 2 different controls: GAPDH and CD30 ( TNFRSF8 ). (B) CSF3R protein expression measured by IHC (cytospin preparations) to compare untreated (left) with ruxolitinib-treated (right) cell lines. The histogram illustrates the pattern of IHC quantification (two-way ANOVA: ∗∗∗ P < .001). ANOVA, analysis of variance.

Article Snippet: Quantitative RT-PCR analysis to quantify the relative level of CSF3R expression was performed using a commercial TaqMan probe with a dye label (FAM) (Hs01114420_m1; Thermo Fisher Scientific).

Techniques: Expressing, Derivative Assay

Journal: Blood Advances

Article Title: JAK/STAT blockade reverses the malignant phenotype of Hodgkin and Reed-Sternberg cells

doi: 10.1182/bloodadvances.2021006336

Figure Lengend Snippet: CSF3R protein expression in primary tumors. (A) Representative examples of CSF3R in HL primary tumors, with strong, faint, and negative IHC expression. (B) mIF for CSF3R (green) and CD30 (red).

Article Snippet: Quantitative RT-PCR analysis to quantify the relative level of CSF3R expression was performed using a commercial TaqMan probe with a dye label (FAM) (Hs01114420_m1; Thermo Fisher Scientific).

Techniques: Expressing