Fig. 3: Reducing FASN protein levels improves ATRA-mediated neutrophil differentiation of APL cells.

A–D NB4 cells were stably transduced with non-targeting shRNA (SHC002) or shRNAs targeting FASN (shFASN_1 and shFASN_2) lentiviral vectors and differentiated with 1 μM ATRA for 1, 2 or 3 days. A FASN western blot analysis of control and shFASN (shFASN_1, _2) expressing NB4 cell populations. B–C NBT reduction in ATRA-treated NB4 control (SHC002) and FASN knockdown (shFASN_1, _2) cells. B Representative images of NBT assays in control and FASN depleted NB4 cells. Scale 20 µM. C Quantification of the percentage of NBT⁺ cells. D Flow cytometry analysis of CD11b surface expression NB4 control (SHC002) and FASN knockdown (shFASN_1, _2) NB4 cells upon ATRA treatment. E–G NB4 cells were treated with the indicated C75 concentrations for 3 days in combination with ATRA. E–F NBT reduction during ATRA-mediated neutrophil differentiation of NB4 control and C75 treated cells. E Representative images of NBT assays in control and C75 treated NB4 cells upon ATRA-mediated differentiation. Scale 20 µM. F Quantification of the percentage of NBT⁺ cells. G Flow cytometry analysis of CD11b surface expression in NB4 control and C75 treated cells upon ATRA-mediated differentiation. H–J NB4 cells were treated with the indicated Orlistat concentrations for 3 days in combination with ATRA. H–I NBT reduction during ATRA-mediated neutrophil differentiation of NB4 control and Orlistat treated cells. H Representative images of NBT assays in control and Orlistat treated NB4 cells upon ATRA-mediated differentiation. Scale 20 µM I Quantification of the percentage of NBT⁺ cells. J Flow cytometry analysis of CD11b surface expression in NB4 control and Orlistat treated cells upon ATRA-mediated differentiation. Data are represented as a mean (n = 3), Error bars: SD.