Figure 3 | Cell Death & Differentiation

Figure 3

From: Cyclin-dependent kinases 7 and 9 specifically regulate neutrophil transcription and their inhibition drives apoptosis to promote resolution of inflammation

Figure 3

Human neutrophils express CDKs and their cyclin binding partners. (a) Whole cell or (b) cytoplasmic/nuclear differential lysates prepared after 4 h incubation in control, R-roscovitine (20 μM), LPS (100 ng/ml) or R-roscovitine and LPS and blotted for CDKs known to be inhibited by R-roscovitine as well as the cyclin binding partners of CDKs 7 and 9. (c) Low levels of CDK2 in human neutrophils confirmed using the proliferating cell lines HL-60 and HepG2 as a positive control. Densitometry on n=3 presented as mean±S.E.M., analysed using ImageJ software. All gels are representative of at least n=3 experiments. Confocal microscopy in neutrophils either unstimulated (d and e) or stimulated with LPS (100 ng/ml) (f and g) and probed for CDK7 (d and f) or 9 (e and g) by indirect immunofluorescence. Panels show transmitted light/bright field (BF), DAPI nuclear staining (blue), and CDK7 or 9 staining (red), final panel shows merged image (MERGE) demonstrating nuclear co-localisation (pink). Size bar represents 5 μm scale. Leica SP confocal microscope × 630 oil immersion. Representative images from at least n=3 experiments

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