Skip to main content
. 2019 Dec 11;14(12):e0225977. doi: 10.1371/journal.pone.0225977

Fig 3. ELANE and CTSG contribute to cell lysis-induced protein degradation in RUNX1-ETO-depleted Kasumi-1 cells.

Fig 3

A) RUNX1-ETO dependent mRNA expression of ELANE and CTSC/K/L/S in Kasumi-1 cells was determined by qRT-PCR and normalized to housekeeping control. Data are shown as mean 2-ΔΔCT +/- SD (n = 3) and p-values were calculated by two-sided student´s t-test (*p<0.05, **p<0.01). B) Whole cell lysates of Kasumi-1/shRE and control cells have been prepared in SDS sample buffer and analyzed for ELANE protein expression by Western Blot. C) Kasumi-1/ctrl and Kasumi-1/shRE cells were consecutively electroporated with ELANE-specific or control siRNA on days five and eight after lentiviral transduction with control virus or shRE. Whole cell lysates were prepared in high-salt lysis buffer on day eleven after RUNX1-ETO depletion and protein expression was analyzed by Western Blot. D) Kasumi-1 cells were transduced with CTSG-specific shRNA (Kasumi-1/shCTSG) or control virus (Kasumi-1/ctrl), followed by transduction with RUNX1-ETO-specific shRNA or control virus, two days after the first lentiviral transduction. Whole cell lysates were prepared in high-salt lysis buffer on day twelve after RUNX1-ETO knockdown (respectively day 14 after CTSG knockdown) and analyzed by Western Blot. Western Blots are representative for three independent experiments.