Journal: Scientific Reports

Article Title: SHOC2 scaffold protein modulates daunorubicin-induced cell death through p53 modulation in lymphoid leukemia cells

doi: 10.1038/s41598-020-72124-1

Figure Lengend Snippet: ( a , b ) Cell viability of Nalm-6 and Reh cells stable transduced with shRNA for SHOC2 or SCR control were evaluated in a 4-day MTT assay after treatment with increasing concentrations of Daunorubicin (10ηM, 25ηM, 50ηM, 100ηM and 150ηM). Results are expressed as mean absorbance [A475 nm–A650 nm] with standard deviation (S.D.) from three independent experiments. Representative viability curves after 72 h of Daunorubicin exposure show how SHOC2 absence has greatly increased daunorubicin resistance (* p < 0.05). ( c , d ) SHOC2 inhibition cleared impaired apoptosis-induced cell death in both Reh and Nalm-6 cells (*** p < 0.001) after Daunorubicin treatment for 72 h (respective IC 50 values). The bar-graphs represent mean with S.D. from three independent experiments. Statistically significant analyses are indicated by asterisks: * p < 0.05, ** p < 0.01.

Article Snippet: The commercial human Leukemic cell lines Nalm-6 (DSMZ ACC128), Reh (ATCC CRL-8286) and Jurkat (ATCC TIB-152) were cultured in RPMI 1640 medium (Corning, Manassas, VA, USA) supplemented with 10% fetal bovine serum (Gibco BRL, Carlsbad, CA, USA), penicillin (100 U/mL) and streptomycin (100 μg/mL) in a humidified atmosphere with 5%CO 2 at 37 °C, as recommended by ATCC.

Techniques: Transduction, shRNA, MTT Assay, Standard Deviation, Inhibition

Journal: Scientific Reports

Article Title: SHOC2 scaffold protein modulates daunorubicin-induced cell death through p53 modulation in lymphoid leukemia cells

doi: 10.1038/s41598-020-72124-1

Figure Lengend Snippet: ( a ) Representative immunoblot of SHOC2 knockdown (obtained from two independent shRNA) significantly impaired ERK1/2 phosphorylation. ( b ) Combined genetic inhibition of SHOC2 with Daunorubicin treatment for 24 h (40ηM) in Nalm-6 cells. Immunoblots reveal reduced p53 expression, impaired Serine 46 phosphorylation and consequently reduced p21 expression upon Daunorubicin-induced damage. Important apoptotic effectors were also affected as observed for p38 phosphorylation inhibition and reduced BAX/BCL-2 ratio (representative bar-graph in the left panel). ( c ) HUWE1 knockdown in Nalm-6 cells greatly increased SHOC2, p53 and p21 expression, confirming SHOC2-p53 interaction. As observed in immunoblot panel in ( b ) after Daunorubicin-induced damage, in SHOC2 absence decreased p53 phosphorylation and also reduced HUWE1 expression.

Article Snippet: The commercial human Leukemic cell lines Nalm-6 (DSMZ ACC128), Reh (ATCC CRL-8286) and Jurkat (ATCC TIB-152) were cultured in RPMI 1640 medium (Corning, Manassas, VA, USA) supplemented with 10% fetal bovine serum (Gibco BRL, Carlsbad, CA, USA), penicillin (100 U/mL) and streptomycin (100 μg/mL) in a humidified atmosphere with 5%CO 2 at 37 °C, as recommended by ATCC.

Techniques: Western Blot, shRNA, Inhibition, Expressing