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Fig. 3 | BMC Molecular and Cell Biology

Fig. 3

From: Lack of Nck1 protein and Nck-CD3 interaction caused the increment of lipid content in Jurkat T cells

Fig. 3

The effects of Nck1 on lipid content in Jurkat T cell. a Deficiency of Nck1 and inhibition of Nck–CD3 interaction caused an increase of lipid mass/cell mass ratio. Cells were unstimulated or stimulated with anti-CD3 antibody (OKT3) alone for 5 min or pre-treated with AX-024 for 30 min followed by OKT3 stimulation for 5 min. Lipid mass/cell mass ratio was monitored by holotomographic microscopy and analysed by Tomostudio software. Data are represented as the mean ± SD from three independent studies, n = 3. b Deletion of Nck1 and inhibition of Nck/CD3 binding was associated with increase lipid droplet. Cells were treated as above and stained with Oil Red O before observing under the inverted microscopy. Image represented one from five separated experiments, n = 5 (top panel). Lipid content in cells was quantified using ImageJ software and presented as mean ± SD of the percentage of Oil Red O-positive area from three independent experiments (button panel). c Re-constituted Nck1-depleted cells (N1KO) with wild-type Nck1 reverted the amount of lipid droplet to normal as Jurkat T cells. N1KO cells were re-constituted with wild-type Nck1 and treated as above before conducting the Oil Red O staining. Picture represented one from three separated experiments, n = 3 (top panel). Lipid droplet in cells was quantified by ImageJ software and presented as mean ± SD of the percentage of Oil Red O-positive area from three independent experiments, n = 3 (button panel). ns = non-significant, **p < 0.01, ***p < 0.001, ****p < 0.0001

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