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HMGN2 controls IL4R/PI3K–AKT signaling and proliferation via R-loops. A - D R-loop detection by S9.6 dot-blot assay. (A, C) Representative images showing R-loop formation in Ly8 and U2932 cells treated with siRNA or overexpressing <t>RNase</t> <t>H</t> (oe-RNase H) to deplete R-loops. (B, D) R-loop intensity quantification in Ly8 and U2932 cells. HMGN2 knockdown reduces R-loop formation, and RNase H further decreases R-loops in HMGN2 knockdown cells. E - F IL4R mRNA expression in Ly8 (E) and U2932 (F) cells. G - H Cell proliferation in both Ly8 (G) and U2932 (H) cells. I - T Western blot analysis of PI3K/AKT signaling proteins. Western blots for IL4R, AKT, p-AKT, PI3K, and p-PI3K in Ly8 (I) and U2932 (Q) cells. Data are presented as means ± SDs. * p < 0.01; ** p < 0.001; *** p < 0.0001
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HMGN2 controls IL4R/PI3K–AKT signaling and proliferation via R-loops. A - D R-loop detection by S9.6 dot-blot assay. (A, C) Representative images showing R-loop formation in Ly8 and U2932 cells treated with siRNA or overexpressing <t>RNase</t> <t>H</t> (oe-RNase H) to deplete R-loops. (B, D) R-loop intensity quantification in Ly8 and U2932 cells. HMGN2 knockdown reduces R-loop formation, and RNase H further decreases R-loops in HMGN2 knockdown cells. E - F IL4R mRNA expression in Ly8 (E) and U2932 (F) cells. G - H Cell proliferation in both Ly8 (G) and U2932 (H) cells. I - T Western blot analysis of PI3K/AKT signaling proteins. Western blots for IL4R, AKT, p-AKT, PI3K, and p-PI3K in Ly8 (I) and U2932 (Q) cells. Data are presented as means ± SDs. * p < 0.01; ** p < 0.001; *** p < 0.0001
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HMGN2 controls IL4R/PI3K–AKT signaling and proliferation via R-loops. A - D R-loop detection by S9.6 dot-blot assay. (A, C) Representative images showing R-loop formation in Ly8 and U2932 cells treated with siRNA or overexpressing <t>RNase</t> <t>H</t> (oe-RNase H) to deplete R-loops. (B, D) R-loop intensity quantification in Ly8 and U2932 cells. HMGN2 knockdown reduces R-loop formation, and RNase H further decreases R-loops in HMGN2 knockdown cells. E - F IL4R mRNA expression in Ly8 (E) and U2932 (F) cells. G - H Cell proliferation in both Ly8 (G) and U2932 (H) cells. I - T Western blot analysis of PI3K/AKT signaling proteins. Western blots for IL4R, AKT, p-AKT, PI3K, and p-PI3K in Ly8 (I) and U2932 (Q) cells. Data are presented as means ± SDs. * p < 0.01; ** p < 0.001; *** p < 0.0001
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HMGN2 controls IL4R/PI3K–AKT signaling and proliferation via R-loops. A - D R-loop detection by S9.6 dot-blot assay. (A, C) Representative images showing R-loop formation in Ly8 and U2932 cells treated with siRNA or overexpressing <t>RNase</t> <t>H</t> (oe-RNase H) to deplete R-loops. (B, D) R-loop intensity quantification in Ly8 and U2932 cells. HMGN2 knockdown reduces R-loop formation, and RNase H further decreases R-loops in HMGN2 knockdown cells. E - F IL4R mRNA expression in Ly8 (E) and U2932 (F) cells. G - H Cell proliferation in both Ly8 (G) and U2932 (H) cells. I - T Western blot analysis of PI3K/AKT signaling proteins. Western blots for IL4R, AKT, p-AKT, PI3K, and p-PI3K in Ly8 (I) and U2932 (Q) cells. Data are presented as means ± SDs. * p < 0.01; ** p < 0.001; *** p < 0.0001
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New England Biolabs rnase h reaction buffer
HMGN2 controls IL4R/PI3K–AKT signaling and proliferation via R-loops. A - D R-loop detection by S9.6 dot-blot assay. (A, C) Representative images showing R-loop formation in Ly8 and U2932 cells treated with siRNA or overexpressing <t>RNase</t> <t>H</t> (oe-RNase H) to deplete R-loops. (B, D) R-loop intensity quantification in Ly8 and U2932 cells. HMGN2 knockdown reduces R-loop formation, and RNase H further decreases R-loops in HMGN2 knockdown cells. E - F IL4R mRNA expression in Ly8 (E) and U2932 (F) cells. G - H Cell proliferation in both Ly8 (G) and U2932 (H) cells. I - T Western blot analysis of PI3K/AKT signaling proteins. Western blots for IL4R, AKT, p-AKT, PI3K, and p-PI3K in Ly8 (I) and U2932 (Q) cells. Data are presented as means ± SDs. * p < 0.01; ** p < 0.001; *** p < 0.0001
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HMGN2 controls IL4R/PI3K–AKT signaling and proliferation via R-loops. A - D R-loop detection by S9.6 dot-blot assay. (A, C) Representative images showing R-loop formation in Ly8 and U2932 cells treated with siRNA or overexpressing <t>RNase</t> <t>H</t> (oe-RNase H) to deplete R-loops. (B, D) R-loop intensity quantification in Ly8 and U2932 cells. HMGN2 knockdown reduces R-loop formation, and RNase H further decreases R-loops in HMGN2 knockdown cells. E - F IL4R mRNA expression in Ly8 (E) and U2932 (F) cells. G - H Cell proliferation in both Ly8 (G) and U2932 (H) cells. I - T Western blot analysis of PI3K/AKT signaling proteins. Western blots for IL4R, AKT, p-AKT, PI3K, and p-PI3K in Ly8 (I) and U2932 (Q) cells. Data are presented as means ± SDs. * p < 0.01; ** p < 0.001; *** p < 0.0001
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HMGN2 controls IL4R/PI3K–AKT signaling and proliferation via R-loops. A - D R-loop detection by S9.6 dot-blot assay. (A, C) Representative images showing R-loop formation in Ly8 and U2932 cells treated with siRNA or overexpressing <t>RNase</t> <t>H</t> (oe-RNase H) to deplete R-loops. (B, D) R-loop intensity quantification in Ly8 and U2932 cells. HMGN2 knockdown reduces R-loop formation, and RNase H further decreases R-loops in HMGN2 knockdown cells. E - F IL4R mRNA expression in Ly8 (E) and U2932 (F) cells. G - H Cell proliferation in both Ly8 (G) and U2932 (H) cells. I - T Western blot analysis of PI3K/AKT signaling proteins. Western blots for IL4R, AKT, p-AKT, PI3K, and p-PI3K in Ly8 (I) and U2932 (Q) cells. Data are presented as means ± SDs. * p < 0.01; ** p < 0.001; *** p < 0.0001
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HMGN2 controls IL4R/PI3K–AKT signaling and proliferation via R-loops. A - D R-loop detection by S9.6 dot-blot assay. (A, C) Representative images showing R-loop formation in Ly8 and U2932 cells treated with siRNA or overexpressing RNase H (oe-RNase H) to deplete R-loops. (B, D) R-loop intensity quantification in Ly8 and U2932 cells. HMGN2 knockdown reduces R-loop formation, and RNase H further decreases R-loops in HMGN2 knockdown cells. E - F IL4R mRNA expression in Ly8 (E) and U2932 (F) cells. G - H Cell proliferation in both Ly8 (G) and U2932 (H) cells. I - T Western blot analysis of PI3K/AKT signaling proteins. Western blots for IL4R, AKT, p-AKT, PI3K, and p-PI3K in Ly8 (I) and U2932 (Q) cells. Data are presented as means ± SDs. * p < 0.01; ** p < 0.001; *** p < 0.0001

Journal: Journal of Translational Medicine

Article Title: HMGN2 deficiency drives DLBCL progression through impaired R-loop formation and PI3K-AKT hyperactivation

doi: 10.1186/s12967-026-08064-7

Figure Lengend Snippet: HMGN2 controls IL4R/PI3K–AKT signaling and proliferation via R-loops. A - D R-loop detection by S9.6 dot-blot assay. (A, C) Representative images showing R-loop formation in Ly8 and U2932 cells treated with siRNA or overexpressing RNase H (oe-RNase H) to deplete R-loops. (B, D) R-loop intensity quantification in Ly8 and U2932 cells. HMGN2 knockdown reduces R-loop formation, and RNase H further decreases R-loops in HMGN2 knockdown cells. E - F IL4R mRNA expression in Ly8 (E) and U2932 (F) cells. G - H Cell proliferation in both Ly8 (G) and U2932 (H) cells. I - T Western blot analysis of PI3K/AKT signaling proteins. Western blots for IL4R, AKT, p-AKT, PI3K, and p-PI3K in Ly8 (I) and U2932 (Q) cells. Data are presented as means ± SDs. * p < 0.01; ** p < 0.001; *** p < 0.0001

Article Snippet: DNA was extracted from Ly8 and U2932 cells transfected with si-HMGN2, oe-HMGN2, or corresponding negative controls (si-NC/oe-NC), followed by overnight incubation at 37 °C in RNase H buffer with or without treatment with RNase H (Beyotime, Shanghai, China).

Techniques: Dot Blot, Knockdown, Expressing, Western Blot