Figure 4.
DCAF13 deficiency impaired T cell proliferation. (A) CFSE signals were detected by flow cytometry to evaluate the proliferative capacity of naïve T cells from CD4 Cre−Dcaf13fl/fl (WT) and CD4 Cre+Dcaf13fl/fl (cKO) mice under anti-CD3/CD28 stimulation. (B) BrdU incorporation was detected by flow cytometry to evaluate DNA replication of naïve T cells from CD4 Cre−Dcaf13fl/fl (WT) and CD4 Cre+Dcaf13fl/fl (cKO) mice under anti-CD3/CD28 stimulation. (C) Flow cytometry analysis of T cell homeostatic proliferation in vivo reconstituted in CD45.1+mice. Naïve T cells isolated from CD45.2+ CD4 Cre−Dcaf13fl/fl or CD4 Cre+Dcaf13fl/fl were labeled with CellTrace Violet and then transferred into CD45.1+ mice as the control and experimental groups, respectively. 4 d after transfer, CellTrace dilution in donor cells from the recipients’ lymph nodes was analyzed by flow cytometry. Representative flow cytometry pictures (left) and corresponding quantitation of the percentage of Celltracelo cells (right; n = 6) are shown. (D) CFSE signals were detected by flow cytometry to evaluate the proliferative capacity of naïve T cells from ERT2 Cre+Dcaf13fl/fl cells under anti-CD3/CD28 stimulation. Tamoxifen-treated T cells were regarded as DCAF13 cKO cells, and those without tamoxifen were regarded as WT controls. The percentage of individual peaks was quantified by FlowJo V10 software. (E) BrdU incorporation was detected by flow cytometry to evaluate DNA replication of naïve T cells from ERT2 Cre+Dcaf13fl/fl mice under anti-CD3/CD28 stimulation. Tamoxifen-treated T cells were regarded as DCAF13 cKO cells, and those without tamoxifen were regarded as WT controls (n = 6). (F) Cell cycle analysis of CD4+ T and CD8+ T cells from ERT2 Cre+Dcaf13fl/fl mice based on the DAPI content. The percentage of individual phase was quantified by FlowJo V10 software. Representative flow cytometry pictures (left) and corresponding quantitation (right; n = 6) are shown. Data are presented as the mean ± SD. A two-tailed Student’s t test was used. ns is not significant. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

DCAF13 deficiency impaired T cell proliferation. (A) CFSE signals were detected by flow cytometry to evaluate the proliferative capacity of naïve T cells from CD4 Cre−Dcaf13fl/fl (WT) and CD4 Cre+Dcaf13fl/fl (cKO) mice under anti-CD3/CD28 stimulation. (B) BrdU incorporation was detected by flow cytometry to evaluate DNA replication of naïve T cells from CD4 Cre−Dcaf13fl/fl (WT) and CD4 Cre+Dcaf13fl/fl (cKO) mice under anti-CD3/CD28 stimulation. (C) Flow cytometry analysis of T cell homeostatic proliferation in vivo reconstituted in CD45.1+mice. Naïve T cells isolated from CD45.2+ CD4 Cre−Dcaf13fl/fl or CD4 Cre+Dcaf13fl/fl were labeled with CellTrace Violet and then transferred into CD45.1+ mice as the control and experimental groups, respectively. 4 d after transfer, CellTrace dilution in donor cells from the recipients’ lymph nodes was analyzed by flow cytometry. Representative flow cytometry pictures (left) and corresponding quantitation of the percentage of Celltracelo cells (right; n = 6) are shown. (D) CFSE signals were detected by flow cytometry to evaluate the proliferative capacity of naïve T cells from ERT2 Cre+Dcaf13fl/fl cells under anti-CD3/CD28 stimulation. Tamoxifen-treated T cells were regarded as DCAF13 cKO cells, and those without tamoxifen were regarded as WT controls. The percentage of individual peaks was quantified by FlowJo V10 software. (E) BrdU incorporation was detected by flow cytometry to evaluate DNA replication of naïve T cells from ERT2 Cre+Dcaf13fl/fl mice under anti-CD3/CD28 stimulation. Tamoxifen-treated T cells were regarded as DCAF13 cKO cells, and those without tamoxifen were regarded as WT controls (n = 6). (F) Cell cycle analysis of CD4+ T and CD8+ T cells from ERT2 Cre+Dcaf13fl/fl mice based on the DAPI content. The percentage of individual phase was quantified by FlowJo V10 software. Representative flow cytometry pictures (left) and corresponding quantitation (right; n = 6) are shown. Data are presented as the mean ± SD. A two-tailed Student’s t test was used. ns is not significant. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal