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( A ) Acute viral infection model. ( B and C ) qRT-PCR analysis (B) and immunoblots (C) of NSD2 expression in CD44 hi CD8 + T cells at different time points postinfection relative to naïve CD8 + T cells [CD44 low CD8; day (d) 0]. β-actin as the loading control. Quantifications and representative histograms were shown ( n = 5). ( D , G , and H ) Representative flow cytometry histograms of <t>GP33</t> + CD8 + T cells in spleens and lymph nodes from WT mice and Nsd2 CKO mice. Quantifications of frequency and absolute cell number at day 8 (D, n = 5 for spleen; n = 9 for lymph nodes), day 15 (G, n = 6), and day 45 (H, n = 8 for spleen; n = 9 for lymph nodes) postinfection were shown. ( E , I , and J ) Representative histograms and quantifications of GP33 + MPECs in spleens and lymph nodes from WT mice and Nsd2 CKO mice at day 8 (E, n = 5 for spleen; n = 9 for lymph nodes from WT mice; n = 6 for lymph nodes from Nsd2 CKO mice), day 15 (I, n = 5 for spleen from WT mice; n = 6 for spleen from Nsd2 CKO mice; n = 6 for lymph nodes), and day 45 (J, n = 6 for spleen; n = 5 for lymph nodes) postinfection were shown. ( F ) Quantifications of viral load in spleen, lung, and liver from WT mice and Nsd2 CKO mice at day 5 postinfection ( n = 9). Data are presented as means ± SEM pooled from two independent experiments (G and H) or from one representative experiment of at least three independent experiments [(B) to (F), (I), and (J)]. Statistical analysis was performed using one-way analysis of variance (ANOVA) [(B) and (C)] and two-tailed unpaired Student’s t test [(D) to (J)]. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.
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( A ) Acute viral infection model. ( B and C ) qRT-PCR analysis (B) and immunoblots (C) of NSD2 expression in CD44 hi CD8 + T cells at different time points postinfection relative to naïve CD8 + T cells [CD44 low CD8; day (d) 0]. β-actin as the loading control. Quantifications and representative histograms were shown ( n = 5). ( D , G , and H ) Representative flow cytometry histograms of <t>GP33</t> + CD8 + T cells in spleens and lymph nodes from WT mice and Nsd2 CKO mice. Quantifications of frequency and absolute cell number at day 8 (D, n = 5 for spleen; n = 9 for lymph nodes), day 15 (G, n = 6), and day 45 (H, n = 8 for spleen; n = 9 for lymph nodes) postinfection were shown. ( E , I , and J ) Representative histograms and quantifications of GP33 + MPECs in spleens and lymph nodes from WT mice and Nsd2 CKO mice at day 8 (E, n = 5 for spleen; n = 9 for lymph nodes from WT mice; n = 6 for lymph nodes from Nsd2 CKO mice), day 15 (I, n = 5 for spleen from WT mice; n = 6 for spleen from Nsd2 CKO mice; n = 6 for lymph nodes), and day 45 (J, n = 6 for spleen; n = 5 for lymph nodes) postinfection were shown. ( F ) Quantifications of viral load in spleen, lung, and liver from WT mice and Nsd2 CKO mice at day 5 postinfection ( n = 9). Data are presented as means ± SEM pooled from two independent experiments (G and H) or from one representative experiment of at least three independent experiments [(B) to (F), (I), and (J)]. Statistical analysis was performed using one-way analysis of variance (ANOVA) [(B) and (C)] and two-tailed unpaired Student’s t test [(D) to (J)]. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.
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( A ) Acute viral infection model. ( B and C ) qRT-PCR analysis (B) and immunoblots (C) of NSD2 expression in CD44 hi CD8 + T cells at different time points postinfection relative to naïve CD8 + T cells [CD44 low CD8; day (d) 0]. β-actin as the loading control. Quantifications and representative histograms were shown ( n = 5). ( D , G , and H ) Representative flow cytometry histograms of <t>GP33</t> + CD8 + T cells in spleens and lymph nodes from WT mice and Nsd2 CKO mice. Quantifications of frequency and absolute cell number at day 8 (D, n = 5 for spleen; n = 9 for lymph nodes), day 15 (G, n = 6), and day 45 (H, n = 8 for spleen; n = 9 for lymph nodes) postinfection were shown. ( E , I , and J ) Representative histograms and quantifications of GP33 + MPECs in spleens and lymph nodes from WT mice and Nsd2 CKO mice at day 8 (E, n = 5 for spleen; n = 9 for lymph nodes from WT mice; n = 6 for lymph nodes from Nsd2 CKO mice), day 15 (I, n = 5 for spleen from WT mice; n = 6 for spleen from Nsd2 CKO mice; n = 6 for lymph nodes), and day 45 (J, n = 6 for spleen; n = 5 for lymph nodes) postinfection were shown. ( F ) Quantifications of viral load in spleen, lung, and liver from WT mice and Nsd2 CKO mice at day 5 postinfection ( n = 9). Data are presented as means ± SEM pooled from two independent experiments (G and H) or from one representative experiment of at least three independent experiments [(B) to (F), (I), and (J)]. Statistical analysis was performed using one-way analysis of variance (ANOVA) [(B) and (C)] and two-tailed unpaired Student’s t test [(D) to (J)]. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.
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( A ) Acute viral infection model. ( B and C ) qRT-PCR analysis (B) and immunoblots (C) of NSD2 expression in CD44 hi CD8 + T cells at different time points postinfection relative to naïve CD8 + T cells [CD44 low CD8; day (d) 0]. β-actin as the loading control. Quantifications and representative histograms were shown ( n = 5). ( D , G , and H ) Representative flow cytometry histograms of <t>GP33</t> + CD8 + T cells in spleens and lymph nodes from WT mice and Nsd2 CKO mice. Quantifications of frequency and absolute cell number at day 8 (D, n = 5 for spleen; n = 9 for lymph nodes), day 15 (G, n = 6), and day 45 (H, n = 8 for spleen; n = 9 for lymph nodes) postinfection were shown. ( E , I , and J ) Representative histograms and quantifications of GP33 + MPECs in spleens and lymph nodes from WT mice and Nsd2 CKO mice at day 8 (E, n = 5 for spleen; n = 9 for lymph nodes from WT mice; n = 6 for lymph nodes from Nsd2 CKO mice), day 15 (I, n = 5 for spleen from WT mice; n = 6 for spleen from Nsd2 CKO mice; n = 6 for lymph nodes), and day 45 (J, n = 6 for spleen; n = 5 for lymph nodes) postinfection were shown. ( F ) Quantifications of viral load in spleen, lung, and liver from WT mice and Nsd2 CKO mice at day 5 postinfection ( n = 9). Data are presented as means ± SEM pooled from two independent experiments (G and H) or from one representative experiment of at least three independent experiments [(B) to (F), (I), and (J)]. Statistical analysis was performed using one-way analysis of variance (ANOVA) [(B) and (C)] and two-tailed unpaired Student’s t test [(D) to (J)]. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.
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Image Search Results


( A ) Acute viral infection model. ( B and C ) qRT-PCR analysis (B) and immunoblots (C) of NSD2 expression in CD44 hi CD8 + T cells at different time points postinfection relative to naïve CD8 + T cells [CD44 low CD8; day (d) 0]. β-actin as the loading control. Quantifications and representative histograms were shown ( n = 5). ( D , G , and H ) Representative flow cytometry histograms of GP33 + CD8 + T cells in spleens and lymph nodes from WT mice and Nsd2 CKO mice. Quantifications of frequency and absolute cell number at day 8 (D, n = 5 for spleen; n = 9 for lymph nodes), day 15 (G, n = 6), and day 45 (H, n = 8 for spleen; n = 9 for lymph nodes) postinfection were shown. ( E , I , and J ) Representative histograms and quantifications of GP33 + MPECs in spleens and lymph nodes from WT mice and Nsd2 CKO mice at day 8 (E, n = 5 for spleen; n = 9 for lymph nodes from WT mice; n = 6 for lymph nodes from Nsd2 CKO mice), day 15 (I, n = 5 for spleen from WT mice; n = 6 for spleen from Nsd2 CKO mice; n = 6 for lymph nodes), and day 45 (J, n = 6 for spleen; n = 5 for lymph nodes) postinfection were shown. ( F ) Quantifications of viral load in spleen, lung, and liver from WT mice and Nsd2 CKO mice at day 5 postinfection ( n = 9). Data are presented as means ± SEM pooled from two independent experiments (G and H) or from one representative experiment of at least three independent experiments [(B) to (F), (I), and (J)]. Statistical analysis was performed using one-way analysis of variance (ANOVA) [(B) and (C)] and two-tailed unpaired Student’s t test [(D) to (J)]. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Journal: Science Advances

Article Title: Histone methyltransferase NSD2 shapes CD8 + T cell fates through the H3K36me2- Zbtb20 axis

doi: 10.1126/sciadv.aee9670

Figure Lengend Snippet: ( A ) Acute viral infection model. ( B and C ) qRT-PCR analysis (B) and immunoblots (C) of NSD2 expression in CD44 hi CD8 + T cells at different time points postinfection relative to naïve CD8 + T cells [CD44 low CD8; day (d) 0]. β-actin as the loading control. Quantifications and representative histograms were shown ( n = 5). ( D , G , and H ) Representative flow cytometry histograms of GP33 + CD8 + T cells in spleens and lymph nodes from WT mice and Nsd2 CKO mice. Quantifications of frequency and absolute cell number at day 8 (D, n = 5 for spleen; n = 9 for lymph nodes), day 15 (G, n = 6), and day 45 (H, n = 8 for spleen; n = 9 for lymph nodes) postinfection were shown. ( E , I , and J ) Representative histograms and quantifications of GP33 + MPECs in spleens and lymph nodes from WT mice and Nsd2 CKO mice at day 8 (E, n = 5 for spleen; n = 9 for lymph nodes from WT mice; n = 6 for lymph nodes from Nsd2 CKO mice), day 15 (I, n = 5 for spleen from WT mice; n = 6 for spleen from Nsd2 CKO mice; n = 6 for lymph nodes), and day 45 (J, n = 6 for spleen; n = 5 for lymph nodes) postinfection were shown. ( F ) Quantifications of viral load in spleen, lung, and liver from WT mice and Nsd2 CKO mice at day 5 postinfection ( n = 9). Data are presented as means ± SEM pooled from two independent experiments (G and H) or from one representative experiment of at least three independent experiments [(B) to (F), (I), and (J)]. Statistical analysis was performed using one-way analysis of variance (ANOVA) [(B) and (C)] and two-tailed unpaired Student’s t test [(D) to (J)]. ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: For intracellular cytokines staining, cells were stimulated with complete T cell medium containing gp33 peptide (200 ng/ml, no. HY-P1569, MedChemExpress), DNase I (0.1 mg/ml, Roche), and GolgiStop Transport Inhibitor (BD Biosciences) at 37°C for 6 hours followed by surface staining, fixing, and permeabilizing using a Cytofix/Cytoperm Fixation/Permeabilization Kit (BD Biosciences) according to the manufacturer’s instructions.

Techniques: Infection, Quantitative RT-PCR, Analysis, Western Blot, Expressing, Control, Flow Cytometry, Two Tailed Test