Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Myeloid-specific HNRNPA2B1 deficiency disrupts macrophage function and in vivo responses.

doi: 10.1093/jimmun/vkaf073

Figure Lengend Snippet: Figure 1. HNRNPA2B1 KO mice show altered immune responses following endotoxic shock in vivo. (A) Temperature change in CTL and HNRNPA2B1 KO mice after i.p. injection with 5 mg/kg LPS. N ¼ 25 CTL, 26 KO. (B–N) Cytokine levels in serum, spleen, and liver of mice treated with 5 mg/kg LPS 6 h postinjection. (B) IFNG_serum, (C) CCL2_serum, (D) CCL3_serum, (E) CSF1_serum, (F) CCL5_serum, (G) IL6_spleen, (H)CCL2_spleen, (I) CCL3_spleen, (J) CSF2_spleen, (K) CXCL1_spleen, (L) IL20_liver, (M) VEGF_liver, (N) TIMP1_liver. Student’s t tests were performed using GraphPad Prism. Asterisks indicate significant differences between mouse lines (P ≤0.05, P ≤0.01).

Article Snippet: Membranes were blocked with PBS, supplemented with 5% (w/v) nonfat dry milk for 1 h, and probed with primary antibody overnight with either HNRNPA2B1 (1:1000, Santa Cruz Biotechnology, sc-374053).

Techniques: In Vivo, Injection

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Myeloid-specific HNRNPA2B1 deficiency disrupts macrophage function and in vivo responses.

doi: 10.1093/jimmun/vkaf073

Figure Lengend Snippet: Figure 2. Macrophage and neutrophil levels are elevated in the HNRNPA2B1 KO mice and show altered macrophage activation following endotoxic shock. (A) Macrophage levels from CTL and HNRNPA2B1 KO mice blood, measured using a flow cytometry panel. (B) Neutrophil levels from CTL and HNRNPA2B1 KO mice blood, measured using a flow cytometry panel. (C) Macrophage levels from CTL and HNRNPA2B1 KO mice spleen, measured using a flow cytometry panel. (D) Neutrophil levels from CTL and HNRNPA2B1 KO mice spleen, measured using a flow cytometry panel. (E) Level of MHCII marker on macrophage surface in the spleen; analysis performed using flow cytometry. (F–H) Level of activation markers on BMDMs ex vivo; analysis performed using flow cytometry: (F) MHCII, (G) CD86, (H) CD80. Student’s t tests were performed using GraphPad Prism. Asterisks indicate significant differences between mouse lines (P ≤0.05, P ≤0.01).

Article Snippet: Membranes were blocked with PBS, supplemented with 5% (w/v) nonfat dry milk for 1 h, and probed with primary antibody overnight with either HNRNPA2B1 (1:1000, Santa Cruz Biotechnology, sc-374053).

Techniques: Activation Assay, Flow Cytometry, Marker, Ex Vivo

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Myeloid-specific HNRNPA2B1 deficiency disrupts macrophage function and in vivo responses.

doi: 10.1093/jimmun/vkaf073

Figure Lengend Snippet: Figure 3. HNRNPA2B1 KO mice are susceptible to Salmonella infection. (A) Survival of CTL and HNRNPA2B1 KO mice after i.p. Salmonella infection. (B) CFU measurement of Salmonella bacterial load in mouse spleen. (C–G) Cytokine levels in serum of CTL and KO mice infected with Salmonella were measured by Elisa, (C) IFNG, (D) IL12 (P70), (E) CXCL5, (F) IL13, (G) CCL11.

Article Snippet: Membranes were blocked with PBS, supplemented with 5% (w/v) nonfat dry milk for 1 h, and probed with primary antibody overnight with either HNRNPA2B1 (1:1000, Santa Cruz Biotechnology, sc-374053).

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Myeloid-specific HNRNPA2B1 deficiency disrupts macrophage function and in vivo responses.

doi: 10.1093/jimmun/vkaf073

Figure Lengend Snippet: Figure 4. HNRNPA2B1 KO macrophages have altered IFN signaling in response to LPS, and KO macrophages fail to clear the pathogen and switch to an alternate form of cell death. (A) Volcano plot of differentially expressed (DE) genes in HNRNPA2B1 KO BMDMs at baseline. (B) Volcano plot of DE genes in HNRNPA2B1 KO BMDMs under LPS stimulation. (C) Gene ontology analysis of downregulated genes at baseline. (D) Gene ontology analysis of upregulated genes at baseline. (E) Gene ontology analysis of downregulated genes post–LPS stimulation. (F) Gene ontology analysis of upregulated genes post–LPS stimulation. (G) Heat map of IFNG response genes’ normalized counts in CTL and KO cells post–LPS stimulation. (H–K) Cytokine levels as measured by ELISA from BMDM supernatant. The supernatant was harvested from CTL and KO cultured BMDMs; and multiplex cytokine analysis was performed for (H) CXCL5 (LIX), (I) CCL22 (MDC), (J) CXCL9 (MIG), (K) IL10. Each dot represents BMDMs from an individual animal. Error bars represent the standard deviation of biological triplicates. Student’s t tests were performed using GraphPad Prism. Asterisks indicate significant differences (P ≤0.05, P ≤0.01). (L) Bacterial growth in ex vivo–infected BMDMs over 8-h time course comparing HNRNPA2B1 KO to Cre CTLs. (I) Rate of total cell death in BMDMs under inflammasome activation. (M) Levels of released IL1b in CTL and KO macrophages under inflammasome activation. (N) Levels of apoptosis markers in BMDMs under inflammasome activation. Student’s t tests were performed using GraphPad Prism. Asterisks indicate significant differences between mouse lines (P ≤0.05, P ≤0.01).

Article Snippet: Membranes were blocked with PBS, supplemented with 5% (w/v) nonfat dry milk for 1 h, and probed with primary antibody overnight with either HNRNPA2B1 (1:1000, Santa Cruz Biotechnology, sc-374053).

Techniques: Enzyme-linked Immunosorbent Assay, Cell Culture, Multiplex Assay, Standard Deviation, Ex Vivo, Infection, Activation Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Myeloid-specific HNRNPA2B1 deficiency disrupts macrophage function and in vivo responses.

doi: 10.1093/jimmun/vkaf073

Figure Lengend Snippet: Figure 5. HNRNPA2B1 regulates IFNG signaling through alternative splicing. (A) CTL BMDM nuclear and cytoplasmic fractions were analyzed using Western blots after treatment with a panel of immune stimuli to assess changes in HNRNPA2B1 localization. (B) Volcano plot of DE genes (black) and common DE and alternatively spliced genes in orange in KO BMDMs. (C) Gene ontology analysis of genes that are differentially expressed as well as alternatively spliced in KO BMDMs. (D–H) Isoform usage levels analyzed by IUTA showing switching in isoform usage between CTL and KO BMDMs in (D) Ifngr1, (E) Oas3, (F) Stat3, (G) Stat1, (H) Irf7. Student’s t tests were performed using GraphPad Prism. Asterisks indicate significant differences between mouse lines (P ≤0.05, P ≤0.01).

Article Snippet: Membranes were blocked with PBS, supplemented with 5% (w/v) nonfat dry milk for 1 h, and probed with primary antibody overnight with either HNRNPA2B1 (1:1000, Santa Cruz Biotechnology, sc-374053).

Techniques: Alternative Splicing, Western Blot

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Myeloid-specific HNRNPA2B1 deficiency disrupts macrophage function and in vivo responses.

doi: 10.1093/jimmun/vkaf073

Figure Lengend Snippet: Figure 6. HNRNPA2B1 regulates alternative splicing of IFN response genes. (A) IFNGRI levels on macrophages in the spleen was measured using flow cytometry. (B) IFNGRI levels on macrophages was measured ex vivo using flow cytometry. (C) IFNGRII levels on macrophages in the spleen was measured using flow cytometry. (D–G) BMDMs from CTL and KO mice were stimulated with IFNG, IFNB, or IFNA for 24 h. Supernatants were removed and ELISAs were performed to measure (D) IL6, (E) MCP1, (F) MIP1b, and (G) RANTES. Student’s t tests were performed using GraphPad Prism. Asterisks indicate significant differences between mouse lines (P ≤0.05, P ≤0.01) (H) Bacterial growth in ex vivo–infected BMDMs over 8 h time course comparing HNRNPA2B1 KO to Cre CTLs. (I) Schematic outlining impacts of HNRNPA2B1 knockout in vivo.

Article Snippet: Membranes were blocked with PBS, supplemented with 5% (w/v) nonfat dry milk for 1 h, and probed with primary antibody overnight with either HNRNPA2B1 (1:1000, Santa Cruz Biotechnology, sc-374053).

Techniques: Alternative Splicing, Flow Cytometry, Ex Vivo, Infection, Knock-Out, In Vivo

Journal: Cell reports

Article Title: A KIF1C-CNBP motor-adaptor complex for trafficking mRNAs to cell protrusions

doi: 10.1016/j.celrep.2025.115346

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: mouse monoclonal anti-hnRNPA2 , Santa Cruz , cat# sc-53531; RRID: AB_2248245.

Techniques: Recombinant, In Vitro, Transfection, Magnetic Beads, Protease Inhibitor, Electron Microscopy, cDNA Synthesis, ISH Cell Assay, In Situ, Negative Control, Control, Software

Journal: Cell reports

Article Title: A KIF1C-CNBP motor-adaptor complex for trafficking mRNAs to cell protrusions

doi: 10.1016/j.celrep.2025.115346

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: The following primary antibodies were used: rabbit polyclonal anti-hnRNPH1 (Bethyl, cat# A300-511A; 1:10,000), rabbit monoclonal anti-hnRNPH2 (Abcam, cat# ab179439; 1:1,000), rabbit polyclonal hnRNPF (Abcam, cat# ab50982; 1:1,000), mouse monoclonal hnRNPA2 (Santa Cruz, cat# sc-53531; 1:1,000), mouse monoclonal anti-CNBP (Proteintech, cat# 67109; 1:3,000), rabbit polyclonal anti-CNBP (ThermoFisher, cat# PA5-35241; 1:1,000), rabbit polyclonal anti-KIF1C (Bethyl, cat# A301-070A; 1:2,000), rabbit polyclonal anti-KIF1C (Proteintech, cat# 12760-1-AP; 1:1,000), mouse monoclonal hnRNPK (Santa Cruz, cat# sc-28380), rabbit monoclonal anti-GAPDH (Proteintech, cat# 2118), rabbit polyclonal anti-GFP (Invitrogen, cat# A-11122; 1:2,000) and rabbit polyclonal anti-Cherry (Abcam, cat# ab183628; 1:5,000).

Techniques: Recombinant, In Vitro, Transfection, Magnetic Beads, Protease Inhibitor, Electron Microscopy, cDNA Synthesis, ISH Cell Assay, In Situ, Negative Control, Control, Software

Journal: Cell reports

Article Title: A KIF1C-CNBP motor-adaptor complex for trafficking mRNAs to cell protrusions

doi: 10.1016/j.celrep.2025.115346

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: mouse monoclonal anti-hnRNPA2 , Santa Cruz , cat# sc-53531; RRID: AB_2248245.

Techniques: Recombinant, In Vitro, Transfection, Magnetic Beads, Protease Inhibitor, Electron Microscopy, cDNA Synthesis, ISH Cell Assay, In Situ, Negative Control, Control, Software