Journal: The Journal of Biological Chemistry

Article Title: Epitope mirroring between the malaria surface proteins PfGARP and PIESP2 identifies a knob-associated complex in infected erythrocytes

doi: 10.1016/j.jbc.2026.113291

Figure Lengend Snippet: Identification and validation of PIESP2 by GM7mAb. A , the codon-optimized PIESP2 construct was synthesized lacking two predicted transmembrane domains and a signal peptide. Nomenclature of defined domains, expression construct, and putative topology are shown. B TRX-PIESP2 protein containing a His-tag, located after TRX protein, was used for affinity purification and fusion protein was purified by Mono Q FPLC. C , Ponceau S-stained image of dose-dependent TRX and TRX-PIESP2, followed by immunoblotting with GM7mAb are shown. D , ELISA-based detection of TRX and TRX-PIESP2 by serial dilution of GM7mAb. E , immunoblotting with GM7mAb using lysates from wild-type 3D7 parasites and PfGARP knockout lines generated at NIH and HMS (only the NIH knockout lane is shown in the left panel ). An anti-Pfaldolase antibody was used as a loading control ( right panel ). F , immunoblot analysis of PfGARP knockout and PIESP2 knockout parasite lysates probed with GM7mAb. The left panel shows the corresponding Ponceau S-stained blot for the immunoblots presented in the right panels . The 48 kDa band was detected in the PfGARP knockout lysate but was absent in the PIESP2 knockout lysate.

Article Snippet: High parasitemia cultures of mature-stage wild-type 3D7, two independent PfGARP knockouts (NIH and HMS), and PIESP2 knockout parasites were purified on Miltenyi Biotech LS columns (MACS) and resuspended in distilled water (1:25 v/v) to lyse the cells.

Techniques: Biomarker Discovery, Construct, Synthesized, Expressing, Affinity Purification, Purification, Staining, Western Blot, Enzyme-linked Immunosorbent Assay, Serial Dilution, Knock-Out, Generated, Control

Journal: The Journal of Biological Chemistry

Article Title: Epitope mirroring between the malaria surface proteins PfGARP and PIESP2 identifies a knob-associated complex in infected erythrocytes

doi: 10.1016/j.jbc.2026.113291

Figure Lengend Snippet: Localization of PIESP2 in P . falciparum infected erythrocytes. A , using GM7mAb, ring stage and mixed trophozoite/schizont stage infected RBCs were tested by immunoblotting for temporal expression of PIESP2. B , multiplex immunoblotting of RIPA-lysed wild-type 3D7 and PfGARP KO (NIH and HMS) parasite lysates. Blots were probed with GM7mAb and rabbit anti-Pfaldolase. Signals were detected by Odyssey CLx using conjugated anti-mouse and anti-rabbit fluorescent secondary antibodies. C , IFA of GM7mAb reactivity was performed in wild-type 3D7, PfGARP knockout (HMS), and PIESP2 knockout erythrocytes. Both iRBCs and uninfected RBC s (data not shown) were fixed and permeabilized with cold methanol (−20 o C for 45 min), labeled with GM7mAb and visualized with Alexa Fluor 488-conjugated anti-mouse IgG. Parasite nuclei were labeled with DAPI. D , five reverse motifs found in PfGARP-M1 and PIESP2 are shown.

Article Snippet: High parasitemia cultures of mature-stage wild-type 3D7, two independent PfGARP knockouts (NIH and HMS), and PIESP2 knockout parasites were purified on Miltenyi Biotech LS columns (MACS) and resuspended in distilled water (1:25 v/v) to lyse the cells.

Techniques: Infection, Western Blot, Expressing, Multiplex Assay, Knock-Out, Labeling

Journal: The Journal of Biological Chemistry

Article Title: Epitope mirroring between the malaria surface proteins PfGARP and PIESP2 identifies a knob-associated complex in infected erythrocytes

doi: 10.1016/j.jbc.2026.113291

Figure Lengend Snippet: PIESP2 is a component of knobs. A–C , immunofluorescence (IFA) colocalization of PIESP2 using GM7mAb with two known knob markers (PfEMP1 and KAHRP) in wild-type 3D7 iRBCs. MSP1 was used as a negative control. Parasite nuclei were stained with DAPI. iRBCs were fixed and permeabilized with cold methanol (−20 o C for 45 min). D , immunoblotting with anti-KAHRP rabbit pAb and anti-PIESP2 (GM7mAb) using uRBC ghosts, iRBC ghosts, and biochemically enriched knobs. PIESP2 is detected in iRBCs and detergent-resistant knobs. E , immunogold transmission electron microscopy of wild-type 3D7 parasite infected erythrocytes with GM7mAb. Signal was detected using colloidal gold-labeled secondary antibody. Green arrowheads denote PIESP2 signal at the tip of knobs. Letter P refers to intracellular parasite and E shows erythrocyte cytoplasm. Scale bars 100 nm.

Article Snippet: High parasitemia cultures of mature-stage wild-type 3D7, two independent PfGARP knockouts (NIH and HMS), and PIESP2 knockout parasites were purified on Miltenyi Biotech LS columns (MACS) and resuspended in distilled water (1:25 v/v) to lyse the cells.

Techniques: Immunofluorescence, Negative Control, Staining, Western Blot, Transmission Assay, Electron Microscopy, Infection, Labeling

Journal: The Journal of Biological Chemistry

Article Title: Epitope mirroring between the malaria surface proteins PfGARP and PIESP2 identifies a knob-associated complex in infected erythrocytes

doi: 10.1016/j.jbc.2026.113291

Figure Lengend Snippet: Localization of PfGARP and PIESP2 on the surface of iRBCs. A , differential reactivity of GM7mAb was measured in non-permeabilized wild-type 3D7, PfGARP knockout, and PIESP2 knockout erythrocytes by immunofluorescence assays (IFA). B , mAb7899 served as a negative control. C and D , a montage of representative IFA images for each genotype is shown in panel C . The image displayed in the second WT 3D7/GM7 mAb panel of A is identical to the image shown in the first WT 3D7 panel of C . To quantify the GM7mAb signal observed in panel A, a larger set of images was analyzed. For quantification, individual infected erythrocytes were manually outlined in ImageJ based on visible fluorescence signal. For each cell, mean fluorescence intensity (MFI) was calculated by subtracting the corresponding background signal. Five representative cells per group were analyzed. Data are presented as mean ± SD. Additional details of IFA quantification are described in the Methods section.

Article Snippet: High parasitemia cultures of mature-stage wild-type 3D7, two independent PfGARP knockouts (NIH and HMS), and PIESP2 knockout parasites were purified on Miltenyi Biotech LS columns (MACS) and resuspended in distilled water (1:25 v/v) to lyse the cells.

Techniques: Knock-Out, Immunofluorescence, Negative Control, Infection, Fluorescence

Journal: The Journal of Biological Chemistry

Article Title: Epitope mirroring between the malaria surface proteins PfGARP and PIESP2 identifies a knob-associated complex in infected erythrocytes

doi: 10.1016/j.jbc.2026.113291

Figure Lengend Snippet: Quantification of GM7mAb reactivity by flow cytometry. Distinct localization of PIESP2 and skeleton binding protein 1. A , flow cytometry was performed on live and unfixed iRBCs from wild-type 3D7, PfGARP knockout, and PIESP2 knockout parasites by GM7mAb to measure surface reactivity. GM7mAb detected robust signal in wild-type 3D7 and PfGARP knockout iRBCs and relatively reduced signal in PIESP2 knockout iRBCs. The numerical values at the right of each panel represent mature parasites. Details of data analysis by FlowJo Software and gating strategies are described in the Methods section. B and C , mAb7899 and mouse IgG3 isotype were used as negative controls. D , immunofluorescence assays (IFA) were used to compare the localization of GM7mAb reactivity (PIESP2) with rabbit anti-SBP1 pAb antibody in wild-type 3D7 and PfGARP knockout iRBCs by confocal microscopy. For confocal microscopy using anti-SBP1 pAb, iRBCs were air-dried and fixed at room temperature with 100% acetone for two minutes (details in ). Representative images of wild-type 3D7 and PfGARP KO parasites subject to co-staining are shown in . The IFA signals were detected by Alexa Fluor 488 anti-mouse secondary antibody and Alexa Fluor 568 anti-rabbit secondary antibody, respectively. Images were acquired on a Zeiss LSM 880 confocal microscope equipped with a 63 × /1.4 NA Plan-Apochromat oil-immersion objective. Scale bar: 2 μm.

Article Snippet: High parasitemia cultures of mature-stage wild-type 3D7, two independent PfGARP knockouts (NIH and HMS), and PIESP2 knockout parasites were purified on Miltenyi Biotech LS columns (MACS) and resuspended in distilled water (1:25 v/v) to lyse the cells.

Techniques: Flow Cytometry, Binding Assay, Knock-Out, Software, Immunofluorescence, Confocal Microscopy, Staining, Microscopy

Journal: The Journal of Biological Chemistry

Article Title: Epitope mirroring between the malaria surface proteins PfGARP and PIESP2 identifies a knob-associated complex in infected erythrocytes

doi: 10.1016/j.jbc.2026.113291

Figure Lengend Snippet: Detection of antibodies in human malaria plasma samples by PIESP2 immunoassay. A , blue color-coded three amino acids denote the boundaries of TRX-PIESP2 construct in the full-length PIESP2 (PF3D7_0501200) sequence. Red color-coded amino acids denote the PIESP2 segment recognized by GM7mAb. Bold RTLAD sequence denotes the location of PEXEL motif. B , mapping of GM7mAb recognition segment within PIESP2-2-2 construct (amin acids 155–228). C , Ponceau-S staining of PIESP2 constructs shown in panel B . D , immunoblotting of PIESP2 constructs (panel C) by GM7mAb. E , alphaFold predicted location of PIESP2-2-2 segment recognized by GM7mAb. F , development of an ELISA for the detection of antibodies against PIESP2. Guided by the presence of reverse motifs in PIESP2, a 25-amino acid peptide (PSP-25) contained within PIESP2-2-2 was synthesized and conjugated to biotin via a flexible linker. The specificity of the peptide was validated by ELISA using GM7mAb and TRX-PIESP2 fusion protein . G , ELISA screens of human plasma samples from a malaria-endemic region in Mali, Africa. Negative controls of PBS and uninfected human serum (H1) were included. The reactivity cutoff value is shown in each ELISA panel. Several plasma samples including 169, 179, 190, 199, 223, 228 were reconfirmed by multiple ELISA screens. Additional details are described in the section.

Article Snippet: High parasitemia cultures of mature-stage wild-type 3D7, two independent PfGARP knockouts (NIH and HMS), and PIESP2 knockout parasites were purified on Miltenyi Biotech LS columns (MACS) and resuspended in distilled water (1:25 v/v) to lyse the cells.

Techniques: Clinical Proteomics, Construct, Sequencing, Staining, Western Blot, Enzyme-linked Immunosorbent Assay, Synthesized

Journal: The Journal of Biological Chemistry

Article Title: Epitope mirroring between the malaria surface proteins PfGARP and PIESP2 identifies a knob-associated complex in infected erythrocytes

doi: 10.1016/j.jbc.2026.113291

Figure Lengend Snippet: Cytoadhesion and schematic representation of knob topology. A , wild-type 3D7, PfGARP knockout, and PIESP2 knockout parasites were evaluated for static binding to human cerebral microvascular endothelial cells (HBEC-5i-CRL-3245 line. ATCC). Bound iRBCs were fixed, stained with Wright-Giemsa, and quantified by microscopy. Effect of GM7mAb, mAb7899, and mouse IgG3 isotype was measured as outlined in the section. Each parasite line was evaluated in a minimum of three independent experiments. B , schematic representation of PIESP2 trafficking to knobs via Maurer’s clefts in infected erythrocytes. C and D , schematic representation of parasite-derived adhesion proteins including PfEMP1, PIESP2, and PfGARP anchored on knobs as putative ligands for host endothelial receptors.

Article Snippet: High parasitemia cultures of mature-stage wild-type 3D7, two independent PfGARP knockouts (NIH and HMS), and PIESP2 knockout parasites were purified on Miltenyi Biotech LS columns (MACS) and resuspended in distilled water (1:25 v/v) to lyse the cells.

Techniques: Knock-Out, Binding Assay, Staining, Microscopy, Infection, Derivative Assay

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: Role of PVPAC-Exo-circEif3c in regulating AF biological functions and its potential mechanism. PVPAC-derived exosomal circEif3c (Exo-circEif3c) promoted AFs migration and proliferation, whereas silencing exosomal circEif3c suppresses these processes. (A) Time-course analysis of circEif3c expression in AFs after Exo-circEif3c treatment (0, 6, and 12 h; 0 h as control). (B) Stable silencing efficiency and specificity of circEif3c in AFs; Exo-siR-control served as the control. (C and D) Effects of PVPAC-Exo-siR- circEif3c-1 and -2 on AF migration and proliferation assessed by wound healing and proliferation assays. Scratch closure percentage and migrated cell numbers were quantified using ImageJ and GraphPad Prism 9.5, scale bar = 150 μm. (E) and (F) FCM analysis of AF proliferation and apoptosis following treatment with PVPAC-Exo-circEif3c, Exo-miR-96–5p, and Ad-MEOX2 interaction. (G) Western blot analysis of vimentin, PHF20L1, and MEOX2 expression in AFs under high glucose and circEif3c modulation. (H) Effects of Exo-circEif3c on the expression of vimentin, PHF20L1, MEOX2, and LC3 in AFs. GAPDH was used as a loading control. All data above are presented as mean ± SD from three independent experiments. vs. the control group, ∗P < 0.05, ∗∗P < 0.01(one-way ANOVA with Dunnett's post-hoc test), n (the number of experiments) = 3.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Derivative Assay, Migration, Expressing, Control, Western Blot

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: The regulatory mchanisms of miR-96-5p in AF biology. (A) Time-course analysis of miR-96–5p expression in AFs treated with PVPAC-Exo at 0, 6, 12, and 24 h. (B) and (C) AFs were transfected with miR-96–5p mimic and NC mimic for 24 h. Then, the migration ability of AFs through migration experiments (B) and EDU assay (C) were evaluated using Image J and GraphPad Prism 9. vs. control mimic, ∗P < 0.05, ∗∗P < 0.01, n (the number of experiments) = 3, scale bar = 150 μm. (D) Correlation analysis between extracellular ncRNA levels in culture supernatant and intracellular expression of PHF20L1 and MEOX2 by RT-qPCR. (E) Bioinformatic prediction identifying PHF20L1 and MEOX2 as potential targets of miR-96–5p. (F) Predicted miR-96–5p binding sites in the 3′UTR of PHF20L1. (G) Luciferase reporter assay displayed that miR-96–5p mimic significantly reduced luciferase activity in the PHF20L1-WT group, but not in the PHF20L1-Mut group (vs. PHF20L1-Mut, ∗∗ P < 0.01), n (the number of experiments) = 3. (H) Bioinformatics analysis indicated the miR-96–5p binding sites in the 3′UTR of MEOX2. (I) Western blot exhibited no significant change in MEOX2 protein levels upon miR-96–5p overexpression. (J) Interaction network among miR-96–5p, circEif3c, PHF20L1, and MEOX2, constructed using GEPIA, ENCORI, miRNet, NDEx, and Cytoscape. (K) Western blot analysis of PHF20L1 and MEOX2 expression in AFs transfected with control-exosome, OE-exosomes, miR-comtrol mimic, miR-96–5p mimic, and UNC1215, respectively. vs. control-exosome, miR-comtrol mimic, ∗ P < 0.05, ∗∗ P < 0.01, n (the number of experiments) = 3. (L) Predicted protein–protein interaction interface between PHF20L1 and MEOX2 using Zdock 3.0.2 and PyMOL 2.5.5. (M) Co-IP experiments confirmed an interaction between PHF20L1 and MEOX2.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Expressing, Transfection, Migration, EdU Assay, Control, Quantitative RT-PCR, Binding Assay, Luciferase, Reporter Assay, Activity Assay, Western Blot, Over Expression, Construct, Co-Immunoprecipitation Assay

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: CircEif3c modulates AF proliferation and migration via the miR-96-5p/PHF20L 1 /MEOX2 axis. (A–C) Cell migration and proliferation assays. AFs were transfected for 24 h with Ad-GFP, siR-circEif3c, miR-96–5p mimic, or siR-MEOX2. Migration (A) and proliferation (B) were quantified (C). (D–F) AFs were co-incubated for 48 h with control mimic, Exo-(siR-)circEif3c mimic, Exo-(siR-)miR-96–5p mimic, PVPAC-exosome (Exo-control), GW4869, or Exo-siR-pAd-MEOX2. Migration (D) and proliferation (E) were assessed (F), scale bar = 150 μm. (G) Cellular fluorescence immunolocalization. nuclei (DAPI, blue), circEif3c (Cy5, red), miR-96–5p (Cy3, orange-yellow), MEOX2 (GFP, green).Scale bar = 30 μm. The above data were presented as mean ± SD. vs. Ad-GFP group, ∗ P < 0.05, ∗∗ P < 0.01, n (the number of experiments) = 3.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: Migration, Transfection, Incubation, Control, Fluorescence

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: Exosomal circEif3c/miR-96-5p/PHF20L1/MEOX2 axis drives vascular remodeling in vivo. (A) Workflow: a stable PVPAC line over-expressing circEif3c supplied exosomes (Exo-Ad-circEif3c, 10 μg/mouse) that were micro-injected into perivascular adipose tissue (PVAT) surrounding the left carotid artery for 4 weeks to initiate remodeling. Subsequently, after the model was established, treatments with (Exo)-Ad-GFP, (Exo)-Ad- circEif3c, (Exo)-Ad-miR-96–5p, and (Exo)-Ad-Meox2 were administered continuously for 2 weeks, respectively. Normal saline (NS) was used as a negative control. (B) Representative H&E-stained cross-sections and concomitant ultrasonography of the common carotid artery. Black scale bars = 50 μm, yellow scale bars = 1 mm, and white scale bars = 0.1 s. (C) Immunohistochemistry. Scale bars = 20 μm. (D) Western blotting. (E) Quantification of protein levels. (F) Tissue localization of Cy5-labeled circEif3c by immunofluorescence, scale bar = 100 μm. (G) Fluorescence intensity quantification. (H) Comparative fluorescence imaging of vascular sections: (H1) Bright-field H&E vs. dark-field GFP before and after Ad-MEOX2 transfection; Scale bars = 50 μm; (H2) DM-remodeling vs MEOX2-intervention groups. Scale bars = 30 μm. (I) Whole-animal in vivo imaging of Cy5 signal. All quantitative data above are presented as mean ± SD. vs. control, ∗ P < 0.01.∗∗ P < 0.01. n (the number of animals) = 6 in each group.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques: In Vivo, Expressing, Injection, Saline, Negative Control, Staining, Immunohistochemistry, Western Blot, Labeling, Immunofluorescence, Fluorescence, Imaging, Transfection, In Vivo Imaging, Control

Journal: Non-coding RNA Research

Article Title: CircEif3c/miR-96–5p/PHF20L1/MEOX2 axis in perivascular preadipocyte exosomes mediates fibroblast dysfunction and vascular remodeling

doi: 10.1016/j.ncrna.2026.01.006

Figure Lengend Snippet: Schematic illustration of the PVPAC-Exo mediated circEif3c/miR-96–5p/PHF20L1/MEOX2 axis regulating vascular remodeling.

Article Snippet: Male wild-type C57BL mice (3–4 weeks old) were purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd., while circEif3c(−/−) and MEOX2(±) conditional knockout mice were generated by Cyagen Biosciences Inc. (Suzhou, China).

Techniques:

Journal: iScience

Article Title: A role for nucleolin in functional improvement in stroke

doi: 10.1016/j.isci.2026.116134

Figure Lengend Snippet: Ischemic cortical stroke increases nucleolin expression in excitatory neurons (A) Representative coronal section stained with DAPI with stroke (red dashed line) and area of interest (yellow box). (B) Representative in situ hybridization images of DAPI (nuclear counterstain), Slc17a7 (excitatory cortical neurons), Gad1 (inhibitory neurons), and nucleolin at time points across acute to chronic stroke. The dashed box indicates the region shown at higher resolution and magnification in accompanying panels. Scale bars, 20 μm, n = 5 animals per time point. (C) Quantification of nucleolin mRNA expression in excitatory cortical neurons post-stroke. Stroke induces both early and chronic expression of nucleolin in Slc17a7 + cells. 1-way ANOVA with post-hoc Tukey correction. Data are represented as mean ± SEM with data points representing individual animals. ∗ p = 0.039, ∗∗∗ p = 0.003 (3D post stroke), p = 0.004 (7D post stroke). (D) Quantification of nucleolin mRNA expression in inhibitory cortical neurons post-stroke. No significant change in nucleolin expression was detected in Gad1 + cells. Data are represented as mean ± SEM with data points representing individual animals. (E) Representative images of nucleolin protein expression in sham stroke and 7 days post-stroke mice. Scale bars, 10 μm, n = 5 animals per group. (F) Quantification of nucleolin protein expression post-stroke. Stroke significantly increases nucleolin expression 7 days post-stroke ( p = 0.0005 via t test). Data are represented as mean ± SEM with data points representing individual animals.

Article Snippet: Nucleolin heterozygous knockout mice , Doron-Mandel et al. , Jackson Laboratories , JAX 037120.

Techniques: Expressing, Staining, In Situ Hybridization

Journal: iScience

Article Title: A role for nucleolin in functional improvement in stroke

doi: 10.1016/j.isci.2026.116134

Figure Lengend Snippet: Heterozygous deletion of the nucleolin GAR domain is detrimental to post-stroke axonal sprouting (A) Representative in situ hybridization images show reduced nucleolin expression in GAR +/- mice. The dashed box indicates the region shown at higher resolution and magnification in accompanying panels. Scale bars, 20 μm, n = 5 animals per group. (B) Quantification of nucleolin mRNA in GAR wild type (GAR +/+ ) and heterozygous deletion (GAR +/- ) mice. GAR +/- mice show a significant reduction in nucleolin mRNA puncta in excitatory cortical neurons ( p = 0.004 via t test). Data are represented as mean ± SEM with data points representing individual animals. (C) Schematic shows the general surgery procedure. Generated via BioRender. (D) Quantitative cortical mapping of peri-infarct axonal sprouting in GAR heterozygous deletion mice compared to wild type littermates. Injection site noted via white circle. Stroke site indicated via black symbol. Heterozygous deletion of the nucleolin GAR domain significantly decreases peri-infarct axonal sprouting when compared to wild-type littermate controls ( p = 0.006 via Hotelling’s T2 test). (E) Axonal puncta distribution within concentric rings centered at the injection origin (0,0). Heterozygous deletion of the nucleolin GAR domain significantly reduces axon detection at distances greater than 3 mm from the injection site (∗ p = 0.0329, ∗∗ p = 0.00286, and ∗∗∗∗ p < 0.0001, general linear mixed model with post-hoc Tukey’s correction). Data are represented as mean ± SEM. (F) Representative cortical flat-map images of wild-type and heterozygous GAR deletion mice 1 month following cortical stroke. White circle indicates the tracer injection site, and the red dashed circle indicates the outer bounds of infarct. (G) Heterozygous deletion of the nucleolin GAR domain results in an approximately 2-fold increase in infarct size one month post stroke ( p = 0.0078 via t test). n = 5 animals per group for all measurements. Data are represented as mean ± SEM with data points representing individual animals.

Article Snippet: Nucleolin heterozygous knockout mice , Doron-Mandel et al. , Jackson Laboratories , JAX 037120.

Techniques: In Situ Hybridization, Expressing, Generated, Injection

Journal: iScience

Article Title: A role for nucleolin in functional improvement in stroke

doi: 10.1016/j.isci.2026.116134

Figure Lengend Snippet: GAR-induced somatic sequestration of nucleolin enhances post-stroke axonal sprouting (A) Quantitative cortical mapping of post-stroke axonal sprouting in flattened mouse cortex ipsilateral to the stroke. Exogenous GAR-induced somatic sequestration of nucleolin, injection site noted by the white circle, results in a significant increase in axonal sprouting to the adjacent somatosensory cortex compared to Dendra2 control virus ( p = 0.02727 via Hotelling’s T2 test). Stroke indicated via a black symbol. (B) Axonal puncta distribution within concentric rings centered at the injection site (0,0). Somatic sequestration of nucleolin significantly increases axon detection locally and distant from the injection site (∗∗ p = 0.00234, ∗∗∗ p = 0.000486, and ∗∗∗∗ p < 0.0001, general linear mixed model with post-hoc Tukey’s correction). Data are represented as mean ± SEM. (C) Representative flatmap images of mice injected with control and GARWT viruses. The white circle indicates the virus injection site, and the red dashed circle indicates the border of the infarct. Yellow square indicates zoomed in image, showing colocalization of Flag and NeuN in virally transduced cells. Scale bars, 20 μm, n = 5–7 animals per group. (D) Quantification of infarct size. Somatic sequestration of nucleolin does not impact infarct size ( p = 0.6989 via t test). n = 7 animals per group for all measurements. Data are represented as mean ± SEM with data points representing individual animals.

Article Snippet: Nucleolin heterozygous knockout mice , Doron-Mandel et al. , Jackson Laboratories , JAX 037120.

Techniques: Injection, Control, Virus

Journal: iScience

Article Title: A role for nucleolin in functional improvement in stroke

doi: 10.1016/j.isci.2026.116134

Figure Lengend Snippet: Nucleolin perturbation enhances post-stroke functional recovery (A) Diagram of grid-walking and pasta matrix tasks, and experimental timeline for task training and assessment post-stroke. (B) Somatic sequestration of nucleolin accelerates recovery in the pasta matrix fine motor control and grid-walking gross motor control tasks. Mice were evaluated for fine motor dexterity as defined by the percentage of baseline pieces of pasta broken. One-week post-stroke, both GARWT + stroke and Control + stroke mice showed a significant deficit in the number of pieces of pasta broken compared to Control alone (∗Control vs. Stroke + Control p = 0.0082, # Control vs. Stroke + GARWT p = 0.0003 via 2-way ANOVA). Three weeks post-stroke, Control + stroke mice continued to show a deficit, whereas GARWT + stroke mice had recovered to a degree with no significant difference from both control groups (∗ Control vs. Control + stroke p = 0.0021, Control vs. GARWT + stroke p = 0.4568, $ Control + stroke vs. GARWT + stroke p = 0.0280 via 2-way ANOVA). By five weeks post-stroke, both groups showed no significant difference from the control groups. N = 8–10 per group. Mice were evaluated for gross motor control as defined by the fold change of baseline percentage right foot faults. One-week post-stroke, Control + stroke and GARWT + stroke mice showed a significant increase in the relative number of foot faults involving the affected limb (∗Control vs. Control + stroke p = 0.0180, # Control vs. GARWT + stroke p = 0.0202 via 2-way ANOVA). At the endpoint of the experiment, 8 weeks post-stroke, Control + stroke mice showed a continued fold change increase in percent foot faults, whereas GARWT + stroke mice were not significantly different from Control alone (∗Control vs. Control + stroke p = 0.0323, Control vs. GARWT + stroke p = 0.3322). n = 12–15 per group. Data are represented as mean ± SEM. (C) Representative DAPI-stained cortical sections of stroke mice treated with control or GARWT virus. Yellow box outlines the peri-infarct region imaged for tissue-level outcomes, red dashed circle indicates infarct bounds. Scale bars, 20 μm. (D) Somatic sequestration of nucleolin has no effect on infarct size or tissue loss. n = 8 per group. p < 0.0001, control vs. control + stroke and control vs. GARWT + stroke via 2-way ANOVA with Tukey’s post-hoc correction. Data are represented as mean ± SEM with data points representing individual animals. (E) Somatic sequestration of nucleolin does not have an effect on Iba1 intensity following cortical stroke. n = 8 per group. Data are represented as mean ± SEM with data points representing individual animals.

Article Snippet: Nucleolin heterozygous knockout mice , Doron-Mandel et al. , Jackson Laboratories , JAX 037120.

Techniques: Functional Assay, Control, Staining, Virus