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ova 323 339  (InvivoGen)
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VPS33B-deficient DCs are impaired in the presentation of MHC class II associated peptides derived from exogenous antigens (A) Representative flow plots of surface 15G4 expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or stimulated with LPS (100 ng/mL) for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % 15G4 + BMDCs and 15G4 MFI (mean fluorescence intensity) to the right. (B) Representative flow plots of surface YAe expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or pulsed with I-E d tetramer (10 μg/mL) for 1 h before being chased for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % YAe + BMDCs to the right. (C) Representative flow plots of CTV-stained OT-II T cells (pre-gated on live, CD4 + , CD8 − ) co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of <t>OVA</t> <t>323-339</t> (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % proliferating (CTV-diluted) OT-II T cells to the right. (D) Representative flow plots of % CD44 + OT-II T cells co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of OVA 323-339 (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % CD44 + OT-II T cells to the right. NP = no peptide (OVA 323-339 ). Error bars shown as mean ± SEM. (A–D) n = 2–4 independent experiments. Statistical analysis was performed by two-way ANOVA. ∗ p < 0 . 01 , ∗∗ p < 0 . 01 , ∗∗∗ p < 0 . 01 , ∗∗∗∗p < 0 . 0001 , n.s. = not significant.
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VPS33B-deficient DCs are impaired in the presentation of MHC class II associated peptides derived from exogenous antigens (A) Representative flow plots of surface 15G4 expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or stimulated with LPS (100 ng/mL) for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % 15G4 + BMDCs and 15G4 MFI (mean fluorescence intensity) to the right. (B) Representative flow plots of surface YAe expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or pulsed with I-E d tetramer (10 μg/mL) for 1 h before being chased for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % YAe + BMDCs to the right. (C) Representative flow plots of CTV-stained OT-II T cells (pre-gated on live, CD4 + , CD8 − ) co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of <t>OVA</t> <t>323-339</t> (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % proliferating (CTV-diluted) OT-II T cells to the right. (D) Representative flow plots of % CD44 + OT-II T cells co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of OVA 323-339 (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % CD44 + OT-II T cells to the right. NP = no peptide (OVA 323-339 ). Error bars shown as mean ± SEM. (A–D) n = 2–4 independent experiments. Statistical analysis was performed by two-way ANOVA. ∗ p < 0 . 01 , ∗∗ p < 0 . 01 , ∗∗∗ p < 0 . 01 , ∗∗∗∗p < 0 . 0001 , n.s. = not significant.
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VPS33B-deficient DCs are impaired in the presentation of MHC class II associated peptides derived from exogenous antigens (A) Representative flow plots of surface 15G4 expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or stimulated with LPS (100 ng/mL) for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % 15G4 + BMDCs and 15G4 MFI (mean fluorescence intensity) to the right. (B) Representative flow plots of surface YAe expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or pulsed with I-E d tetramer (10 μg/mL) for 1 h before being chased for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % YAe + BMDCs to the right. (C) Representative flow plots of CTV-stained OT-II T cells (pre-gated on live, CD4 + , CD8 − ) co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of <t>OVA</t> <t>323-339</t> (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % proliferating (CTV-diluted) OT-II T cells to the right. (D) Representative flow plots of % CD44 + OT-II T cells co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of OVA 323-339 (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % CD44 + OT-II T cells to the right. NP = no peptide (OVA 323-339 ). Error bars shown as mean ± SEM. (A–D) n = 2–4 independent experiments. Statistical analysis was performed by two-way ANOVA. ∗ p < 0 . 01 , ∗∗ p < 0 . 01 , ∗∗∗ p < 0 . 01 , ∗∗∗∗p < 0 . 0001 , n.s. = not significant.
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VPS33B-deficient DCs are impaired in the presentation of MHC class II associated peptides derived from exogenous antigens (A) Representative flow plots of surface 15G4 expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or stimulated with LPS (100 ng/mL) for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % 15G4 + BMDCs and 15G4 MFI (mean fluorescence intensity) to the right. (B) Representative flow plots of surface YAe expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or pulsed with I-E d tetramer (10 μg/mL) for 1 h before being chased for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % YAe + BMDCs to the right. (C) Representative flow plots of CTV-stained OT-II T cells (pre-gated on live, CD4 + , CD8 − ) co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of <t>OVA</t> <t>323-339</t> (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % proliferating (CTV-diluted) OT-II T cells to the right. (D) Representative flow plots of % CD44 + OT-II T cells co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of OVA 323-339 (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % CD44 + OT-II T cells to the right. NP = no peptide (OVA 323-339 ). Error bars shown as mean ± SEM. (A–D) n = 2–4 independent experiments. Statistical analysis was performed by two-way ANOVA. ∗ p < 0 . 01 , ∗∗ p < 0 . 01 , ∗∗∗ p < 0 . 01 , ∗∗∗∗p < 0 . 0001 , n.s. = not significant.
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VPS33B-deficient DCs are impaired in the presentation of MHC class II associated peptides derived from exogenous antigens (A) Representative flow plots of surface 15G4 expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or stimulated with LPS (100 ng/mL) for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % 15G4 + BMDCs and 15G4 MFI (mean fluorescence intensity) to the right. (B) Representative flow plots of surface YAe expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or pulsed with I-E d tetramer (10 μg/mL) for 1 h before being chased for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % YAe + BMDCs to the right. (C) Representative flow plots of CTV-stained OT-II T cells (pre-gated on live, CD4 + , CD8 − ) co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of OVA 323-339 (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % proliferating (CTV-diluted) OT-II T cells to the right. (D) Representative flow plots of % CD44 + OT-II T cells co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of OVA 323-339 (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % CD44 + OT-II T cells to the right. NP = no peptide (OVA 323-339 ). Error bars shown as mean ± SEM. (A–D) n = 2–4 independent experiments. Statistical analysis was performed by two-way ANOVA. ∗ p < 0 . 01 , ∗∗ p < 0 . 01 , ∗∗∗ p < 0 . 01 , ∗∗∗∗p < 0 . 0001 , n.s. = not significant.

Journal: iScience

Article Title: VPS33B regulates MHC class II antigen presentation in dendritic cells to drive CD4 T cell immunity

doi: 10.1016/j.isci.2026.116052

Figure Lengend Snippet: VPS33B-deficient DCs are impaired in the presentation of MHC class II associated peptides derived from exogenous antigens (A) Representative flow plots of surface 15G4 expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or stimulated with LPS (100 ng/mL) for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % 15G4 + BMDCs and 15G4 MFI (mean fluorescence intensity) to the right. (B) Representative flow plots of surface YAe expression on VPS33B WT and VPS33B ΔCSF1R BMDCs left US or pulsed with I-E d tetramer (10 μg/mL) for 1 h before being chased for the indicated time points. Plots were pre-gated on live, CD11b + , CD11c + cells. Graphical quantification of % YAe + BMDCs to the right. (C) Representative flow plots of CTV-stained OT-II T cells (pre-gated on live, CD4 + , CD8 − ) co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of OVA 323-339 (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % proliferating (CTV-diluted) OT-II T cells to the right. (D) Representative flow plots of % CD44 + OT-II T cells co-cultured with VPS33B WT and VPS33B ΔCSF1R sDCs in the presence or absence of OVA 323-339 (0.01 and 0.1 μM) and LPS (100 ng/mL) for 72 h. Graphical quantification of % CD44 + OT-II T cells to the right. NP = no peptide (OVA 323-339 ). Error bars shown as mean ± SEM. (A–D) n = 2–4 independent experiments. Statistical analysis was performed by two-way ANOVA. ∗ p < 0 . 01 , ∗∗ p < 0 . 01 , ∗∗∗ p < 0 . 01 , ∗∗∗∗p < 0 . 0001 , n.s. = not significant.

Article Snippet: OVA 323-339 , Invivogen , Cat#vac-isq.

Techniques: Derivative Assay, Expressing, Fluorescence, Staining, Cell Culture