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Fig. 1 | Journal of Hematology & Oncology

Fig. 1

From: Inhibition of LIN28B impairs leukemia cell growth and metabolism in acute myeloid leukemia

Fig. 1

The effect of silencing LIN28B in AML. a Lentiviral LIN28B shRNA 1, 2, 3, 4, 5 and Scramble-shRNA were transduced in TF-1a cells. Proteins of the knockdown cell lines were harvested at 24, 48, and 90 h time points for Western blot analysis. β-actin was used as a loading control. b RNAs of shRNA 3 and 5 transduced cells were extracted after 1 week of drug selection. qRT-PCR was performed to compare LIN28B transcription level. The expression of LIN28B in each sample was normalized with GAPDH, respectively (n = 3, mean ± SD). *p < 0.01. c Cell viability assay of TF-1a cells transduced with Scramble shRNA or LIN28B shRNA 3, 5. Eight repeats of 10,000 cells in 100 μl of medium each per sample were cultured at various time points: 0, 24, 48, and 96 h by CTG assays. Measurements taken from 24 h onwards were normalized with their respective values obtained at 0 h (n = 8, mean ± SD). n.s. not significant; *p < 0.05; **p < 0.01. d qRT-PCR and Western blot analysis validation of overexpression of LIN28B in TF-1 and HEK293T cells, followed by cell viability assays at 0, 24, and 48 h times. Measurements taken from 24 h onwards were normalized with their respective values obtained at 0 h (n = 8, mean ± SD). n.s. not significant; *p < 0.05; **p < 0.01. e Colony-forming assay of TF1-pEGFP and TF1-LIN28B. The experiments were duplicated and representative pictures were presented (left panel). qRT-PCR analysis confirmed decreased LIN28B expression in LIN28B shRNA3, 5 treated THP-1 cells (middle panel). Quantification of colonies of indicated cell lines (n = 3, mean ± SD). The colony number of LIN28B-shRNAs expression THP-1 cells was significantly reduced than those of Scramble shRNA expressing cells (right panel, *p < 0.01). f Flow cytometric detection of the CD34+CD38− population in pairs of LIN28B knockdown and overexpression cell lines. The representative FACS images were shown in the left panel and bar graphs of the percentages of CD34+CD38− cells were listed in the right panel (n = 2, mean ± SD) (*p < 0.05, **p < 0.01)

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