Fig 2: Ikzf2 transferred into the nucleus of ICOS+ Th cells and bound to the promoter icos gene after S. japonicum infection. (A) Five to 6 weeks after S. japonicum infection, splenocytes were isolated and stained by different fluorescence-labeled antibodies. CD4+ ICOS+ T cells and CD4+ ICOS− T cells were sorted by FACS, respectively. After the separation of nucleoplasmic proteins, the distribution of Ikzf2 was detected by Western blotting. Tubulin and histone H3 were used as cytoplasm and nucleus controls. (B) CD4+ T cells were stimulated with CD3 and CD28 mAb for 24 h, and ICOS+ / − Th cells were selected by magnetic bead sorting. The expression of ICOS was detected by flow cytometry. (C) The DNA fragments of the sorted cells treated with the chromatin shearing kit were analyzed by DNA electrophoresis. The sheared length of DNA should be about 150–900 bp. (D) ChIP results were obtained by qRT-PCR. Anti-Ikzf2 mAb, positive control histone H3 rabbit mAb, and negative control normal rabbit IgG were added to pull down the promoter of ICOS, respectively. Percent Input = 2% × 2(C[T] 2%Input Sample − C[T] IP Sample). Data were shown as three samples in each group from one representative experiment, and experiments were repeated three times with similar results. (E) HEK293T cells were transfected with ICOS promoter reporter plasmids containing the firefly luciferase and the renilla luciferase gene. Simultaneously, 500 ng of Ikzf2-flag plasmids and empty vectors were transfected into HEK293T cells. Twenty-four hours after transfection, IKZF2 expression were detected by Western blot. β-Actin was used as an internal reference protein. (F) The remaining cell lysates were used for the DLR test. The ratio of firefly luciferase activity to Renilla luciferase activity was converted to relative luciferase activity. The means are from three independent experiments and the error bars are SD. **P < 0.01, and ns, not significant (P > 0.05).

Fig 3: Previously reported molecular glue IKZF2 degraders and new molecular glue degraders of IKZF2.a Chemical structures of representative, previously reported molecular glue IKZF2 degraders. b Chemical structures of new IKZF2 degraders and IKZF2 degradation data. Data are presented as mean of n = 2 biological replicates. Source data are provided as a Source Data file.

Fig 4: PK/PD evaluation of PVTX-405 in immune-competent and gene-edited C57BL/6 mice harboring humanized CrbnI391V gene.CrbnI391V mice were administered with vehicle control or PVTX-405 via oral gavage using 100% PEG200 as the dosing vehicle. Mice were sacrificed at indicated time-points, and blood samples, spleen tissues and thymus tissues were harvested for analysis. At each time point, 4 mice were euthanized for each dosing group. a IKZF2 degradation induced by PVTX-405 in spleen and thymus tissues in mice. A method of one-way ANOVA was used for P value calculation: “**”P < 0.01; “****”P < 0.0001. The p values for all PVTX-405 treated groups compared with their corresponding vehicle control are <0.0001, except a p value of 0.0088 for the group (10 mg/kg PVTX-405, Thymus, 24 h) compared with the vehicle control of thymus tissue. Data presented as mean ± SEM, with n = 4 animals for each treatment group. b Concentrations of PVTX-405 in plasma, spleen and thymus at 6, 12, and 24 h time-points and 10, 30, and 100 mg/kg doses. “ND”: not determined. Data presented as mean ± SEM, with n = 4 animals for each treatment group. Source data are provided as a Source Data file.

Fig 5: X-ray crystal structure of IZKF2:CRBN:compound 5 ternary complex.a Ternary structure of IKZF2 Zinc finger 2 (ZF2) domain (green) with CRBN Thalidomide-binding domain (TBD, gray) and compound 5 (yellow sticks) at 1.7 Å resolution (PDB ID: 9DOM). b Detailed interactions of IKZF2:CRBN:Compound 5 ternary structure with hydrogen bonds shown in dashed lines.