Fig. 3

SHQ1 depletion impairs T-ALL cell survival while it spares normal bone marrow cells. a HPB-ALL and patient-derived T-ALL (#1) cells were infected with lentiviruses expressing control (Ctrl, blue) or SHQ1 shRNA (sh1-red or sh2-purple) with GFP as an expression marker. SHQ1 mRNA levels were analyzed in GFP⁺ cells. b Live GFP⁺ cells were counted at the indicated time points and cell growth was plotted as shown. Cell counts of those expressing control shRNA, SHQ1 shRNA-1, or -2 are marked as blue square, red circle, or purple diamond. c Percentages of GFP⁺ cell populations were analyzed by flow cytometry at the indicated time points post infection. d, e Apoptotic cell death was analyzed by Annexin V-PI staining 6 days post infection and quantified in e (blue: control shRNA, purple: SHQ1 shRNA-2), along with immunoblots of SHQ1. Long exposure of SHQ1 bands in bone marrow cells is also shown (Long exp). f Cell viability was assessed by MTT assay in two T-ALL cell lines (HPB-ALL, blue and KOPTK1, green), two primary T-ALL cells (#1, red and #3, purple), and two normal human BM cells (#1, black and #2, brown) expressing either control or SHQ1 shRNA. Absorbance readings of SHQ1-deficient cells relative to control cells were plotted as shown. Data shown represent the means (±SEM) of three biological replicates; *p < 0.05, **p < 0.01, ***p < 0.001, unpaired t-test (a, b, e)