k562 nuclear cell extracts  (Santa Cruz Biotechnology)

Journal: Cold Spring Harbor Molecular Case Studies

Article Title: 1q21.1 deletion and a rare functional polymorphism in siblings with thrombocytopenia-absent radius–like phenotypes

doi: 10.1101/mcs.a004564

Figure Lengend Snippet: Electromobility shift assays (EMSAs) suggest that rs61746197 A > G is a functional SNP. ( A ) Ten micrograms of unstimulated nuclear extract from K562 cells was incubated with reference (Ref, R) or alternative (Alt, A) probe as indicated. Cold Ref probe titrates signal away from Alt at a lower stoichiometric ratio than the converse (arrow and compare lanes 11 – 13 with 6 – 8 ). ( B ) Unstimulated extract ( left panel, U) exhibits a markedly stronger baseline shift than TPA stimulated extract ( right panel, S). Unstimulated nuclear extract (U) when incubated with labeled alternative probe (A) produces a novel secondary shift not observed with either the reference probe (R) or the TPA stimulated extract (S). Addition of RelA–p65 antibody abrogates this secondary shift (arrow).

Article Snippet: EMSAs were performed using TPA-stimulated and unstimulated nuclear extracts of K562 cells (Active Motif).

Techniques: Functional Assay, Incubation, Labeling

Journal: Scientific Reports

Article Title: Down syndrome-associated haematopoiesis abnormalities created by chromosome transfer and genome editing technologies

doi: 10.1038/srep06136

Figure Lengend Snippet: (a) Sequence analyses of GATA1s-ES cells. One allele had an 8-bp deletion and the other had a 17-bp deletion; both resulted in a TGA stop codon in exon 2 of GATA1 . The GATA1 nucleotide (upper line) and amino acid (lower line) sequences are shown for WT-ES and GATA1s-ES cells. An asterisk shows the stop codon. (b) Western blot analyses of erythroid lineage-differentiated cells derived from WT-ES, WT-ES sublines, Ts21-ES lines, GATA1s-ES, and GATA1s/Ts21-ES lines. Anti-BACH1 was used to detect the gene-dosage effect on hChr.21. Anti-GATA1 recognised the C-terminus of both GATA1 and GATA1s protein. Anti-α-tubulin was used as an internal control. HEL cell lysate and K562 nuclear extract were used as positive controls. 10T1/2 whole cell lysate was used as a negative control. Cropped blots were used in this figure. Original full-length blots are shown in . (c, d) Haematopoietic differentiation analyses using WT-ES, WT-ES sublines, Ts21-ES lines, GATA1s-ES, and GATA1s/Ts21-ES lines. Data are the means of 3 independent experiments (±S.D.). The percentage of CD34+, CD41a+/CD42b+, and CD71+/CD235+ cells are shown in each differentiation stage (ES-sac (day 14), megakaryocyte (day 20), and erythroid (day 20)) (c). Statistical analyses were performed by comparison with WT-ES cells (WT-ES1, WT-ES1-1 and WT-ES1-2). *p < 0.05, **p < 0.01 by two-tailed Student's t test. The percentage of CD34-/CD41a-, CD34+/CD41a+ and CD34-/CD41a+ cells are shown in the megakaryocyte stage (d).

Article Snippet: HEL 92.1.7 whole cell lysate and K562 nuclear extract were used as positive controls (sc-2130 and sc-2277, respectively; Santa Cruz Biotechnology).

Techniques: Sequencing, Western Blot, Derivative Assay, Control, Negative Control, Comparison, Two Tailed Test