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. Author manuscript; available in PMC: 2017 Sep 8.
Published in final edited form as: Blood Adv. 2017 Aug 8;1(18):1358–1367. doi: 10.1182/bloodadvances.2017008383

Figure 4. NELF depletion leads to a genome-wide loss of transcription in progenitor cells.

Figure 4

(A) Western blot of NELF-B and -E in human CD34+ HSPCs transfected with a scramble siRNA (ctrl), or siRNA targeting NELF-B or -E. Protein extraction was done on day 3 post-transfection. GAPDH is used as a loading control.

(B) Quantification of western blot in A by imageJ. Protein level is normalized to GAPDH and presented as fold change relative to control samples (n=3, mean ± SEM, **p<0.01).

(C) Metagene analysis showing Pol II occupancy measured by GRO-seq on both sense and anti-sense strands in human CD34+ HSPCs transfected with control-siRNA (ctrl) or siRNA targeting NELF-E (NE-KD). Cells were collected on day 3 post-transfection.

(D) Boxplot analysis to compare pausing index between cells transfected with a control-siRNA (ctrl), or siRNA targeting NELF-B (NB-KD) or –E (NE-KD).

(E) Cumulative distribution function analysis to compare PI distribution in cells transfected with a control-siRNA (ctrl), or siRNA targeting NELF-E (NE-KD)

(F) Venn Diagram showing the overlap of upregulated (“up”) and downregulated genes (“down”) between NELF-B and NELF-E knockdown cells.