Figure 1.

Significantly decreased WIPI-1 mRNA expression in primary AML patient samples and increase in WIPI-1 expression upon ATRA treatment of NB4 APL cells was detected. (a) WIPI-1 mRNA levels of granulocytes from healthy donors and AML patient samples were measured using qPCR. Data represent log2 expression levels and were normalized to the expression levels of the two housekeeping genes HMBS and ABL. For better readability, we multiplied the results by (−1) and excluded Ct values higher than 40 (ΔCt=40−Ct^WIPI-1−(Mean Ct^HMBS and Ct^ABL1) × (−1)). (b) mRNA levels of WIPI-1 expression was measured in control, and ATRA-treated NB4, NB4-R2 APL cell lines at day 4 using qPCR. Values were normalized to the housekeeping gene HMBS and given as n-fold mRNA expression relative to the control of day 4. (c, d) Increase in endogenous WIPI-1 puncta was detected using confocal microscopy and was further quantified using ImageJ software. (e) PI3P:WIPI-1 binding assay of cell lysates from NB4 cells treated with 1 μM ATRA for 6 days. Each membrane was loaded with equal total protein lysate. M.W.U, *P<0.05. **P<0.01, ***P<0.001. M.W.U, Mann–Whitney U-test