Journal: PLOS One

Article Title: KIF11 promotes rheumatoid arthritis pathogenesis by activating M1 macrophage polarization and promoting inflammatory cytokine secretion

doi: 10.1371/journal.pone.0347313

Figure Lengend Snippet: (A) A petal plot demonstrated 672 overlapping DEGs identified from five RA synovial tissue datasets ( GSE55457 , GSE55235 , GSE2053 , GSE12021 , and GSE1919 ). DEGs were selected with thresholds of |log2FC| ≥ 0.5 and adjusted p-value < 0.05. (B) The mRNA expression levels of KIF11 in rheumatoid arthritis (RA) and normal control (NC) synovial tissues were analyzed in six independent GEO datasets: GSE1919 , GSE12021 , GSE55235 , GSE55457 , GSE77298 , and GSE2053 . Data are presented as mean ± SD. ***P < 0.001, compared with NC controls. (C) MH7A cells were treated with TNF-α (10 ng/mL) and IL-1β (10 ng/mL) for 24 h, and the expression of KIF11 was determined via western blotting. (D) The relative expression of KIF11 was shown in histogram. ***P < 0.01, compared with untreated group.

Article Snippet: KIF11 human shRNA lentiviral particles (Locus ID 3832, Catalog Number: TL311921V) and their corresponding control lentiviral particles (Catalog Number: TR30021V) were obtained from Origene Inc.

Techniques: Expressing, Control, Western Blot

Journal: PLOS One

Article Title: KIF11 promotes rheumatoid arthritis pathogenesis by activating M1 macrophage polarization and promoting inflammatory cytokine secretion

doi: 10.1371/journal.pone.0347313

Figure Lengend Snippet: (A) The expression of KIF11 was detected by western blotting. (C) Cell viability was measured by MTT assay at 24, 48, and 72 h post-infection with control shRNA (shCtrl) or KIF11 shRNA (shKIF11) lentivirus. n = 3. ***P < 0.001 vs. Ctrl. shRNA group. (D) Edu incorporation assay. Fluorescence imaging results at 24 and 48 hours. E. The proportion of positively stained cells is displayed in a bar graph, ***P < 0.001. F. Colony formation assay. Representative images (E) and quantification (F) of colony formation assay in Ctrl. shRNA- and KIF11 shRNA infected MH7A cells after two-week culture. ***P < 0.001 vs. Ctrl.shRNA group, ###p < 0.001 vs.untreated cells .

Article Snippet: KIF11 human shRNA lentiviral particles (Locus ID 3832, Catalog Number: TL311921V) and their corresponding control lentiviral particles (Catalog Number: TR30021V) were obtained from Origene Inc.

Techniques: Expressing, Western Blot, MTT Assay, Infection, Control, shRNA, Fluorescence, Imaging, Staining, Colony Assay

Journal: PLOS One

Article Title: KIF11 promotes rheumatoid arthritis pathogenesis by activating M1 macrophage polarization and promoting inflammatory cytokine secretion

doi: 10.1371/journal.pone.0347313

Figure Lengend Snippet: (A) Transwell migration assay of shCtrl- and shKIF11-infected MH7A cells at 24 h and 48 h. Representative images (40 × magnification) and migrated cell quantification (right). Data are mean ± SD ( n = 3). *P < 0.05, **P < 0.01 vs. CtrlshRNA group. (B) The expression of IL-1β, IL-6, and IL-8 was detected by ELISA assay in cell supernatant. ***P < 0.001. (C) Rescue of KIF11 expression in knockdown cells. MH7A cells with stable KIF11 knockdown (shKIF11) were transfected with either a human KIF11 cDNA overexpression plasmid (KIF11-OE) or the empty control vector (Pcmv3 vector). After 24 hours, KIF11 protein expression was analyzed by Western blotting. β-actin served as a loading control. (D) The relative KIF11 protein levels were shown in histogram. Data are presented as mean ± SD (n = 3). ***P < 0.001 vs. the vector-transfected shKIF11 group. (E) Rescue of pro-inflammatory cytokine secretion. The cell culture supernatants from the experiment described in (D) were collected, and the concentrations of TNF-α, IL-1β, IL-6, and IL-8 were measured by ELISA. Data are presented as mean ± SD (n = 3). ***p < 0.001 vs. the control vector-transfected shKIF11 group.

Article Snippet: KIF11 human shRNA lentiviral particles (Locus ID 3832, Catalog Number: TL311921V) and their corresponding control lentiviral particles (Catalog Number: TR30021V) were obtained from Origene Inc.

Techniques: Transwell Migration Assay, Infection, Expressing, Enzyme-linked Immunosorbent Assay, Knockdown, Transfection, Over Expression, Plasmid Preparation, Control, Western Blot, Cell Culture

Journal: PLOS One

Article Title: KIF11 promotes rheumatoid arthritis pathogenesis by activating M1 macrophage polarization and promoting inflammatory cytokine secretion

doi: 10.1371/journal.pone.0347313

Figure Lengend Snippet: (A) KIF11 knockdown inhibited NF-κB p65 phosphorylation. MH7A cells were infected with control shRNA (shCtrl) or KIF11 shRNA (shKIF11) lentivirus for 24 hours. Whole-cell lysates were subjected to Western blotting using antibodies against phospho-p65 (p-p65), total p65, and KIF11. β-actin served as a loading control. (B) The histogram shows the relative ratio of p-p65 to total p65. Data are presented as mean ± SD (n = 3). ***P < 0.001 vs. shCtrl group. (C) KIF11 knockdown impaired nuclear translocation of NF-κB p65. Cytoplasmic and nuclear fractions were extracted from shCtrl- and shKIF11-infected MH7A cells. The distribution of p65 in the cytoplasm (Cyto) and nucleus (Nuc) was analyzed by Western blotting. Lamin B1 and α-tubulin were used as markers for nuclear and cytoplasmic fractions, respectively. (D) The relative abundance of p65 in the nuclear and cytoplasmic fractions were shown in histograms, normalized to their respective loading controls. Data are presented as mean ± SD (n = 3). ***P < 0.001 vs. shCtrl group.

Article Snippet: KIF11 human shRNA lentiviral particles (Locus ID 3832, Catalog Number: TL311921V) and their corresponding control lentiviral particles (Catalog Number: TR30021V) were obtained from Origene Inc.

Techniques: Knockdown, Phospho-proteomics, Infection, Control, shRNA, Western Blot, Translocation Assay

Journal: PLOS One

Article Title: KIF11 promotes rheumatoid arthritis pathogenesis by activating M1 macrophage polarization and promoting inflammatory cytokine secretion

doi: 10.1371/journal.pone.0347313

Figure Lengend Snippet: A. KIF11 was knock down in M1 macrophages for 24 h. B. The histogram of gray value was shown IN Ctrl. shRNA or KIF11 shRNA lentiviruses infected M1 macrophages. **P < 0.01. Interference with KIF11 in M1 macrophages decreased the expression of CD86. The intensity of surface molecules CD14 and CD86 (C) and CD86(D) were determined by flow cytometry in THP-1-derived M0 and M1 cells, shKIF11 lentivirus or ctrl. shRNA lentivirus infected M1 macrophages. E. The histogram of CD86 and CD80 expression was shown. ***P < 0.001.

Article Snippet: KIF11 human shRNA lentiviral particles (Locus ID 3832, Catalog Number: TL311921V) and their corresponding control lentiviral particles (Catalog Number: TR30021V) were obtained from Origene Inc.

Techniques: Knockdown, shRNA, Infection, Expressing, Flow Cytometry, Derivative Assay

Journal: Nature Communications

Article Title: KIF11 prevents retinal endothelial ferroptosis in familial exudative vitreoretinopathy by inhibiting phosphorylation-driven PRDX1 phase separation

doi: 10.1038/s41467-026-71009-7

Figure Lengend Snippet: a Schematic diagram of the integrative bulk RNA-seq analysis in HRECs depleted with CTNNB1 , FZD4 , LRP5 , or TSPAN12 , using the same batch of control (CTRL) cells. Created in BioRender. Li, S. (2026) https://BioRender.com/b75j366 . b Differential gene expression analysis showing the upregulated (red) and downregulated (blue) genes in CTNNB1 , FZD4 , LRP5 , or TSPAN12 knockdown (KD) HRECs. c Heatmap of representative β-catenin downstream targets and knockdown genes. d Heatmap of WGCNA module-trait relationships, where rows represent modules and columns represent HRECs transduced with CTRL, CTNNB1 , FZD4 , LRP5 , or TSPAN12 shRNA. e Heatmap of genes from blue and dark gray modules across all samples. KEGG pathway enrichment of biological process signalings for genes within blue ( f ) and dark gray ( g ) modules. h Hub genes identified by Cytohubba plug-in (MCC, Maximal Clique Centrality) within the dark gray module. i Differential expression of KIF11 in CTRL versus CTNNB1 KD, FZD4 KD, LRP5 KD, or TSPAN12 KD HRECs by bulk RNA-seq, with Log 2 FC of −1.62, −0.92, −1.22, and −1.23, respectively. Box plots show the median (center line), 25th–75th percentiles (bounds of box), and whiskers extending to the minima and maxima values. Relative mRNA levels of KIF11 , CTNNB1 , FZD4 , LRP5 , and TSPAN12 in CTRL versus CTNNB1 KD ( j ), FZD4 KD ( k ), LRP5 KD ( l ), or TSPAN12 KD ( m ) HRECs. n = 3. Western blot analyses and relative quantification of protein levels of KIF11 together with CTNNB1, FZD4, LRP5, or TSPAN12 in CTRL versus CTNNB1 KD ( n ) FZD4 KD ( o ), LRP5 KD ( p ) or TSPAN12 KD ( q ) HRECs, respectively. n = 3. Data are presented as mean ± SD. n represents independent biological replicates. Statistical significance was determined using a two-tailed Student’s t test ( i – q ). Source data are provided as a file. MW molecular weight.

Article Snippet: For rescue experiments, lentiviruses (20 μl at a titer of 1 × 10 9 TU/mL) expressing KIF11, wild-type PRDX1, mutant PRDX1, or an empty vector (control) were obtained from Genechem Co. (Shanghai, China).

Techniques: RNA Sequencing, Control, Gene Expression, Knockdown, Transduction, shRNA, Quantitative Proteomics, Western Blot, Two Tailed Test, Molecular Weight

Journal: Nature Communications

Article Title: KIF11 prevents retinal endothelial ferroptosis in familial exudative vitreoretinopathy by inhibiting phosphorylation-driven PRDX1 phase separation

doi: 10.1038/s41467-026-71009-7

Figure Lengend Snippet: a Representative images of retinal flat mounts from P6 wild-type (WT), Ctnnb1 conditional knockout (cKO) mice, and Tspan12 knockout (KO) mice. Circles indicate vessel outgrowth of the control. Green, Isolectin GS-IB4. Scale bars, 500 μm. b Schematic diagram of scRNA-seq from isolated retinal vascular cells and publicly available datasets. Created in BioRender. Li, S. (2026) https://BioRender.com/b75j366 . c UMAP diagram showing distinct cell clusters of retinas. d Heatmap of the relative Kif11 , Apcdd1 , and Plvap expression within endothelial cells (ECs) across samples. e UMAP diagram of EC subpopulations. f UMAP plot of Kif11 and Mki67 expression. g Stacked bar plot depicting the composition of endothelial cell subpopulations in postnatal and adult groups. h Violin plots showing Kif11 expression in proliferative ECs. The outer violin outline represents the kernel probability density. The embedded box plots indicate the median (center line) and mean (dashed line), the 25th and 75th percentiles (bounds of the box), and the whiskers extending to the minima and maxima. n = 134, 355, 491, and 66 cells per group. i Western blot and relative quantification of KIF11 in retinas from different time points. n = 3. j Pie charts showing annotation of ATAC-seq peak distributions. k ATAC-seq peak signals of Kif11 and Axin2 in isolated cerebellar ECs from control and EC-specific β-catenin-stabilized mice. l Heatmap of ChIP-seq signal intensity around transcription start sites (TSS). m Genome-wide distribution of ChIP-seq peaks. n ChIP-seq peak signals of Kif11 and Axin2 . o Western blot and relative quantification of WT and mutant KIF11 protein levels. n = 3. Representative images ( p ) and quantification of retinal vascular progression ( q ) and vessel density ( r ) from WT and P7 Ctnnb1 cKO mice treated with CTRL or KIF11-overexpressing lentivirus. Circles indicate vessel outgrowth of the KIF11-overexpressed mice. Red, Isolectin GS-IB4. Scale bars, 500 μm. n = 8. Data are presented as mean ± SD. n represents independent biological replicates or the number of mice per group. Statistical significance was determined using two-tailed Student’s t test ( h ), one-way ANOVA with Dunnett’s ( i, o ), or Tukey’s ( q, r ) multiple comparisons test. Source data are provided as a file.

Article Snippet: For rescue experiments, lentiviruses (20 μl at a titer of 1 × 10 9 TU/mL) expressing KIF11, wild-type PRDX1, mutant PRDX1, or an empty vector (control) were obtained from Genechem Co. (Shanghai, China).

Techniques: Knock-Out, Control, Isolation, Expressing, Western Blot, Quantitative Proteomics, ChIP-sequencing, Genome Wide, Mutagenesis, Two Tailed Test

Journal: Nature Communications

Article Title: KIF11 prevents retinal endothelial ferroptosis in familial exudative vitreoretinopathy by inhibiting phosphorylation-driven PRDX1 phase separation

doi: 10.1038/s41467-026-71009-7

Figure Lengend Snippet: a Representative cell index values of CTRL and KIF11 -depleted HRECs. n = 3. b Representative transmission electron microscopy (TEM) images of CTRL and KIF11 -depleted HRECs. Green arrows, autophagosome; red arrows, mitochondria. Scale bars, 500 nm. c Volcano plot of differentially expressed proteins in CTRL and KIF11 KD HRECs. d Bubble plot of the KEGG pathway enrichment analysis for differentially expressed proteins upon KIF11 knockdown. e Heatmap of the altered ferroptosis-related genes in CTRL and KIF11 -depleted HRECs. f Nine-quadrant diagram integrating transcriptomic and proteomic data of CTRL and KIF11 -depleted HRECs, with log 2 FoldChange thresholds of −1 and 1 for transcriptome, and −0.585 and 0.585 for proteome. g Number of genes in each quadrant. h Relative quantification of GPX4 mRNA and protein level in control and KIF11 -depleted HRECs profiled by transcriptome and Proteome. n = 3. Western blot ( i ) and relative quantification ( j ) of ferroptosis-related proteins in control and KIF11 -depleted HRECs. n = 3. k Quantification of relative GPx activity in control and KIF11 -depleted HRECs. n = 3. Representative images ( l ) and relative mean intensity quantification ( m ) of FerroOrange in CTRL and KIF11 -depleted HRECs. Orange, FerroOrange; blue, Hoechst. Scale bars, 10 μm. n = 6. n Relative MDA levels in CTRL and KIF11 -depleted HRECs. n = 3. Representative time-lapse images of BODIPY (red) ( o ) and quantification of BODIPY + cells ( p ) in CTRL and KIF11 -depleted HRECs. Scale bars, 70 μm. n = 3. q Representative images of CTRL and KIF11 -depleted HRECs stained with CellROX and Hoechst. Red, CellROX; blue, Hoechst. Scale bars, 10 μm. r Quantification of relative mean CellROX fluorescence intensity in CTRL and KIF11 -depleted HRECs. n = 7. s Heatmap of phospholipid peroxidation in CTRL and KIF11 -depleted HRECs. Data are presented as mean ± SD. n represents independent biological replicates. Statistical significance was determined using two-tailed Student’s t test ( h, j, k, n ), two-tailed Welch’s t-test ( r ), Mann–Whitney U test ( m ), or two-way ANOVA with Sidak’s multiple comparisons test ( p ). Source data are provided as a file.

Article Snippet: For rescue experiments, lentiviruses (20 μl at a titer of 1 × 10 9 TU/mL) expressing KIF11, wild-type PRDX1, mutant PRDX1, or an empty vector (control) were obtained from Genechem Co. (Shanghai, China).

Techniques: Transmission Assay, Electron Microscopy, Knockdown, Quantitative Proteomics, Control, Western Blot, Activity Assay, Staining, Fluorescence, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: KIF11 prevents retinal endothelial ferroptosis in familial exudative vitreoretinopathy by inhibiting phosphorylation-driven PRDX1 phase separation

doi: 10.1038/s41467-026-71009-7

Figure Lengend Snippet: a Representative cell index values of KIF11 -depleted HRECs treated with Fer-1 or DMSO. n = 3. Representative immunofluorescence images ( b ) and quantification of CellROX ( c ) and FerroOrange ( d ) intensities in KIF11 -depleted HRECs treated with Fer-1 or DMSO. Red, CellROX; orange, FerroOrange; blue, Hoechst. Scale bars, 10 μm. n = 7 ( c ) and 6 ( d ). GPx activity ( e ) and MDA levels ( f ) in HRECs treated with Fer-1 or DMSO. n = 3. Immunoblotting ( g ) and quantification ( h–k ) of core ferroptosis-related proteins in HRECs treated with Fer-1 or DMSO. n = 3. Representative images ( l ) and quantification ( m ) of BODIPY (red), Annexin V (blue), and PI (red) staining in HRECs treated with DMSO or Fer-1. n = 3. n Schematic diagram of immunoprecipitation-mass spectrometry analysis. Created in BioRender. Li, S. (2026) https://BioRender.com/b75j366 . o Top nine potential KIF11-binding partners identified by mass spectrometry. Co-IP analysis of interactions between KIF11-FLAG and IK or PRDX1 ( p ), and between mutant KIF11-FLAG and PRDX1-HA ( q ). r Representative images of the PRDX1 puncta in CTRL and KIF11 -depleted HRECs. Dotted boxes: zoomed-in areas. Arrows, PRDX1 puncta. Red, PRDX1; blue, DAPI. Scale bars, 10 μm. s Quantification of PRDX1 puncta numbers per cell. n = 5. Representative time-lapse FRAP images ( t ), mean percentage recovery of PRDX1-EYFP puncta ( u ), and quantification of PRDX1-EYFP puncta sizes ( v ) in CTRL and KIF11 -depleted HEK293T cells transfected with PRDX1-EYFP. Arrows, the bleached puncta. Green, EYPF; blue, Hoechst. Scale bars, 2 μm. n = 3 ( u ). n = 39 (CTRL) and n = 91 ( KIF11 KD) cells pooled from three independent biological preparations ( v ). The error bar center represents the mean ( v ). w Representative images of PRDX1-EYFP overexpressing CTRL and KIF11 -depleted HEK293T cells before and after 10% 1,6-HD treatment. Green, EYFP; blue, Hoechst. Scale bars, 5 μm. Non-reducing SDS PAGE immunoblot ( x ) and quantification ( y ) of the HWM- and LWM- PRDX1. n = 3. Data are presented as mean ± SD. Unless otherwise specified, n represents independent biological replicates. Statistical significance was determined using two-tailed Student’s t test ( y ), two-tailed Welch’s t-test ( c, f, s ), Mann–Whitney U test ( d, v ), one-way ANOVA with Tukey’s multiple comparisons test (m), or two-way ANOVA with Tukey’s multiple comparisons test ( e, h – k ). Source data are provided as a file.

Article Snippet: For rescue experiments, lentiviruses (20 μl at a titer of 1 × 10 9 TU/mL) expressing KIF11, wild-type PRDX1, mutant PRDX1, or an empty vector (control) were obtained from Genechem Co. (Shanghai, China).

Techniques: Immunofluorescence, Activity Assay, Western Blot, Staining, Immunoprecipitation, Mass Spectrometry, Binding Assay, Co-Immunoprecipitation Assay, Mutagenesis, Transfection, SDS Page, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: KIF11 prevents retinal endothelial ferroptosis in familial exudative vitreoretinopathy by inhibiting phosphorylation-driven PRDX1 phase separation

doi: 10.1038/s41467-026-71009-7

Figure Lengend Snippet: Western blot analysis ( a ) and quantification ( b ) of phosphorylated PRDX1-Tyr194 and KIF11 expression levels. n = 3. Western blot analysis ( c ) and quantification ( d ) of phosphorylated PRDX1-Tyr194 expression levels in HEK293T cells expressing WT or Y194Q-mutant PRDX1. n = 3. e Representative images of PRDX1 in WT or Y194Q-mutant PRDX1-overexpressed HRECs. Dotted boxes, zoomed-in regions. Red, PRDX1; blue, DAPI. Scale bars, 10 μm. f Quantification of PRDX1 puncta numbers per cell in HRECs overexpressing with WT or Y194Q-mutant PRDX1. n = 6. Non-reducing SDS PAGE immunoblot ( g ) and quantification ( h ) of HWM- and LWM- PRDX1 levels in WT and Y194Q-mutant PRDX1 overexpressed HRECs. n = 3. i Prediction of intrinsically disordered regions (IDRs) of WT and Y194Q-mutant PRDX1 using ESpritz ( http://old.protein.bio.unipd.it/espritz/ ). Western blotting ( j ) and quantification ( k ) of HWM- and LWM-PRDX1 in KIF11 KD HRECs treated with larotinib (100 μM) or DMSO. n = 3. Co-IP assays showing the interaction between KIF11-FLAG and either wild-type or Y194Q-mutant PRDX1 ( l ), and relative quantification of PRDX1 binding ( m ). n = 3. Western blotting ( n ) and quantification ( o ) of HWM- and LWM-PRDX1 in KIF11 KD HRECs treated with Src kinase inhibitor PP1 (100 μM) or DMSO. n = 3. p Schematic diagram of Co-IP assays of Src-miniTurboID-FLAG and PRDX1-HA in the presence or absence of KIF11. Created in BioRender. Li, S. (2026) https://BioRender.com/b75j366 . Co-IP assays of Src-miniTurboID-FLAG and PRDX1-HA ( q ), and quantification of streptavidin-bound biotinlated-PRDX1-HA ( r ). n = 3. s Co-IP assays of streptavidin-bound biotinylated-PRDX1-HA by Src-miniTurboID-FLAG. Quantification of streptavidin-pulled PRDX1 (normalized to Src) ( t ), and levels of P-PRDX1 (normalized to PRDX1) in total lysates (INPUT) ( u ), in the presence or absence of overexpressed KIF11-FLAG. n = 3. Data are presented as mean ± SD. n represents independent biological replicates. Statistical significance was determined using two-tailed Student’s t test ( b, d, h, k, m, o, r, u ) or two-tailed Welch’s t-test ( f, t ). Source data are provided as a file.

Article Snippet: For rescue experiments, lentiviruses (20 μl at a titer of 1 × 10 9 TU/mL) expressing KIF11, wild-type PRDX1, mutant PRDX1, or an empty vector (control) were obtained from Genechem Co. (Shanghai, China).

Techniques: Western Blot, Expressing, Mutagenesis, SDS Page, Co-Immunoprecipitation Assay, Quantitative Proteomics, Binding Assay, Two Tailed Test

Journal: Nature Communications

Article Title: KIF11 prevents retinal endothelial ferroptosis in familial exudative vitreoretinopathy by inhibiting phosphorylation-driven PRDX1 phase separation

doi: 10.1038/s41467-026-71009-7

Figure Lengend Snippet: Western blot ( a ) and quantification of PRDX1 protein ( b ) and mRNA levels ( c ) upon PRDX1 KD. n = 3. Representative cell index values ( d ), EdU images ( e ), and quantification ( f ) of EdU + cells (%) in PRDX1 -depleted HRECs. Red, EdU; blue, DAPI. Scale bars, 50 μm. n = 4. g TEM images in CTRL and PRDX1 -depleted HRECs. Green arrows, autophagosome; red arrows, mitochondria. Scale bars, 500 nm. Representative FerroOrange images ( h ) and intensity quantification ( i ) in CTRL and PRDX1 -depleted HRECs. Orange, FerroOrange; blue, Hoechst. Scale bars, 10 μm. n = 6. Relative GPx activity ( n = 3) ( j ), CellROX immunofluorescence images ( k ) and mean intensity quantification ( n = 6) ( l ), and MDA levels ( n = 3) ( m ) in CTRL and PRDX1 KD HRECs. Red, CellROX; blue, Hoechst. Scale bars, 10 μm. n Volcano plots of DEGs upon PRDX1 KD. 3D PCoA plot summarizing bulk RNA-seq sample distribution of CTRL, KIF11 , and PRDX1 KD HRECs ( o ), and of CTRL and all knockdown groups ( p ). q Quantification of ferroptosis-related gene mRNAs in CTRL and PRDX1 -depleted HRECs. n = 3. Western blot ( r ) and quantification ( s ) of ferroptosis-related proteins upon PRDX1 -depleted. n = 3. t Representative cell index values of PRDX1 -depleted HRECs treated with Fer-1 or DMSO. n = 3. Representative immunofluorescence ( u ) and quantification of CellROX ( v , left) and FerroOrange ( v , right) in PRDX1 -depleted HRECs treated with Fer-1 or DMSO. Red, CellROX; orange, FerroOrange; blue, Hoechst. Scale bars, 10 μm. n = 6. GPx activity ( w ) and MDA levels ( x ) in PRDX1 -depleted HRECs treated with Fer-1 or DMSO. n = 3. Immunoblotting ( y ) and quantification ( z ) of core ferroptosis-related proteins in CTRL and PRDX1 -depleted HRECs treated with Fer-1 or DMSO. n = 3. Data are presented as mean ± SD. n represents independent biological replicates. Statistical significance was determined using two-tailed Student’s t test ( j, m, q, s, x ), two-tailed Welch’s t-test ( b, c, l, v ), Mann–Whitney U test ( f, i ), or two-way ANOVA with Tukey’s multiple comparisons test ( w, z ). Source data are provided as a file.

Article Snippet: For rescue experiments, lentiviruses (20 μl at a titer of 1 × 10 9 TU/mL) expressing KIF11, wild-type PRDX1, mutant PRDX1, or an empty vector (control) were obtained from Genechem Co. (Shanghai, China).

Techniques: Western Blot, Activity Assay, Immunofluorescence, RNA Sequencing, Knockdown, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: KIF11 prevents retinal endothelial ferroptosis in familial exudative vitreoretinopathy by inhibiting phosphorylation-driven PRDX1 phase separation

doi: 10.1038/s41467-026-71009-7

Figure Lengend Snippet: a Representative images of retinal flat mounts from P7 littermate control and Kif11 cKO mice. The circles indicate vessel outgrowth of the control. Red, Isolectin GS-IB4. Dotted boxes indicate magnified areas. Scale bars, 500 μm and 50 μm. Quantification of retinal vascular progression ( b ) and vessel density ( c ) of P7 littermate control and Kif11 cKO mice. n = 8. d Representative images of superficial and deep retinal vasculature from P10 littermate control and Kif11 cKO mice. Red, Isolectin GS-IB4. Scale bars, 500 μm. e Representative TEM images of retinas from P7 littermate control and Kif11 cKO mice. Green arrows, autophagosome; red arrows, mitochondria. Scale bars, 500 nm. Western blot analysis ( f ) and quantification ( g ) of phosphorylated PRDX1-Tyr194 in lung lysates from P15 littermate control and Kif11 cKO mice. n = 3. Immunoblotting analysis ( h ) and quantification of relative expression levels ( i ) of core autophagy-related genes in lung lysates from P15 littermate control and Kif11 cKO mice. n = 3. Immunoblotting analysis ( j ) and quantification of relative expression levels ( k ) of core ferroptosis-related genes in lung lysates from P15 littermate control and Kif11 cKO mice. n = 3, Data are presented as mean ± SD. n represents independent biological replicates or the number of mice per group. Statistical significance was determined using a two-tailed Student’s t test ( b, c, g, i, k ). Source data are provided as a file.

Article Snippet: For rescue experiments, lentiviruses (20 μl at a titer of 1 × 10 9 TU/mL) expressing KIF11, wild-type PRDX1, mutant PRDX1, or an empty vector (control) were obtained from Genechem Co. (Shanghai, China).

Techniques: Control, Western Blot, Expressing, Two Tailed Test

Journal: Nature Communications

Article Title: KIF11 prevents retinal endothelial ferroptosis in familial exudative vitreoretinopathy by inhibiting phosphorylation-driven PRDX1 phase separation

doi: 10.1038/s41467-026-71009-7

Figure Lengend Snippet: a Schematic diagram of the Fer-1 treatment design. Created in BioRender. Li, S. (2026) https://BioRender.com/b75j366 . Representative retinal flat mounts ( b ) and quantification of retinal vascular progression ( c ) and vessel density ( d ) in P7 WT and Kif11 cKO mice treated with DMSO, and Kif11 cKO mice treated with Fer-1 (30 mg/kg). Circles indicate vessel outgrowth in Kif11 cKO mice treated with Fer-1. Red, Isolectin GS-IB4. Scale bars, 500 μm. n = 8. Representative retinal flat mounts ( e ) quantification of retinal vascular progression ( f ) and vessel density ( g ) in P7 WT and Ctnnb1 cKO mice treated with DMSO, and Ctnnb1 cKO mice treated with Fer-1 (30 mg/kg). Circles indicate vessel outgrowth in Ctnnb1 cKO mice treated with Fer-1. Red, Isolectin GS-IB4. Scale bars, 500 μm. n = 8. h Representative cell index values of CTRL, WT-, or Y194Q-mutant PRDX1 overexpressing HRECs. n = 3. i Schematic diagram of the rescue design of PRDX1-expressing lentivirus. Created in BioRender. Li, S. (2026) https://BioRender.com/b75j366 . Representative retinal flat mounts ( j ) and quantification of retinal vascular progression ( k ) and vessel density ( l ) in P7 Kif11 cKO mice treated with LV-CTRL, LV-PRDX1-WT, or LV-PRDX1-MUT. Circles indicate vessel outgrowth in the Kif11 cKO mice treated with LV-PRDX1-MUT. Red, Isolectin GS-IB4. Scale bars, 500 μm. n = 8. m Schematic of FEVR pathogenesis driven by β-catenin/KIF11 axis dysfunction. In normal retinal development, Norrin/β-catenin signaling induces KIF11, which competitively binds PRDX1 to inhibit its Src-mediated phosphorylation and subsequent liquid-liquid phase separation (LLPS). Conversely, FEVR-associated mutations in this pathway trigger the loss of KIF11 function and KIF11-PRDX1 interaction, leading to aberrant PRDX1 phosphorylation and LLPS. This protein condensation triggers ferroptosis, ultimately impairing retinal vascularization. Created in BioRender. Li, S. (2026) https://BioRender.com/b75j366 . Data are presented as mean ± SD. n represents independent biological replicates or the number of mice per group. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test ( c, d, f, g, k, l ). Source data are provided as a file.

Article Snippet: For rescue experiments, lentiviruses (20 μl at a titer of 1 × 10 9 TU/mL) expressing KIF11, wild-type PRDX1, mutant PRDX1, or an empty vector (control) were obtained from Genechem Co. (Shanghai, China).

Techniques: Mutagenesis, Expressing, Phospho-proteomics

Journal: Nature Communications

Article Title: Intellectual disability-causing mutations in KIF11 impair microtubule dynamics and dendritic arborization

doi: 10.1038/s41467-026-70522-z

Figure Lengend Snippet: A Schema of selected Microcephaly with or without chorioretinopathy, lymphedema, or intellectual disabilities (MCLID) patient mutations ( Hs: Homo sapiens ) and the corresponding mouse homolog (Mm: Mus Musculus ) (adapted from Schlögel et al. ) on the KIF11 protein. B Experimental timeline. C Confocal projection images of primary hippocampal mouse neurons transfected with control or KIF11 constructs, with the soma in the center of the image. Scale Bar=25 µm. D Soma size quantification of ( C ). N = 20,22,22,19 neurons for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Tukey’s test. E Quantification of dendritic morphology changes using Sholl analysis. N = 17,20,20,16 neurons for NC-GFP, KIF11-OE, KIF11 Y81F, and KIF11 ΔCterm , respectively. Two-way ANOVA followed by Tukey’s test. F Plus-end-out EB3-comet flux in KIF11 dendrites in comparison to NC-GFP. N = 17,17,18,21 dendrites for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. G . Minus-end-out EB3-comets flux in KIF11 dendrites in comparison to NC-GFP. N = 14,16,18,21 dendrites for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. H Percentage of Minus-end-out EB3-comets in KIF11 dendrites in comparison to NC-GFP. N = 17,17,18,22 dendrites for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. I . Length of plus-end-out MT growth for KIF11 dendrites in comparison to NC-GFP. N = 17(248),17(59),18(114),22(184) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. J Length of minus-end-out MT growth for KIF11 dendrites compared to NC-GFP. N = 17(70),17(42),18(27),22(45) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. K Plus-end-out MT growth-rate based on EB3-comet velocities in KIF11 dendrites in comparison to NC-GFP. N = 17(254),17(56),18(130),22(189) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. L Minus-end-out MT growth rate based on EB3-comet velocities in KIF11 dendrites in comparison to NC-GFP. N = 17(59),17(49),18(32),22(41) dendrites (# of comets) for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm , respectively. One-way ANOVA followed by Dunnett’s test. For all graphs ( D–L ), error bars represent ±SEM. P values are listed above respective comparisons. P values are listed above respective comparisons. Source data are provided as a file.

Article Snippet: DIV16 primary hippocampal neurons culture (in 35 mm Mattek dishes) were transfected with 0.5 μg of NC-GFP (Origene TR30013) or KIF11-OE (Genecoepia CS-Mm03465-Lv236-01) for Supplementary Fig. ; or 0.5 μg NC-GFP (Origene TR30013), KIF11-OE (Genecoepia CS-Mm03465-Lv236-01), or Kif11 Y81F (Genecoepia CS-Mm03465-Lv236-02) for Supplementary Fig. . At DIV18, neuronal cultures were washed with PBS and then fixed in a freshly prepared solution of 4% paraformaldehyde for 15 min. After three or more rinses in PBS, the cells were then incubated in 10% normal horse serum(NHS) (Gibco) in PBS-T with 0.1% Triton X-100 for 1 h at room temperature to reduce the non-specific binding of the primary antibody.

Techniques: Transfection, Control, Construct, Comparison

Journal: Nature Communications

Article Title: Intellectual disability-causing mutations in KIF11 impair microtubule dynamics and dendritic arborization

doi: 10.1038/s41467-026-70522-z

Figure Lengend Snippet: A Timeline of experimental design to record miniature excitatory post-synaptic potential (mEPSCs) in mouse primary hippocampal culture expressing NC-GFP or KIF11 constructs. Tetrodotoxin (TTX) was added to ensure mEPSCs and not spontaneous EPSCs were captured. B Two representative traces of mEPSCs for NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm 24–48 h post-transfection. Bar graph of mEPSC amplitude ( C ) and frequency ( D ) in NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm neurons. N = 10,13,12,13 NC-GFP, KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm neurons respectively. One-way ANOVA followed by Tukey’s Multiple comparisons test. Cumulative probability graphs showing no change in mEPSC amplitude ( E ), but reduced frequency ( F ) in KIF11-OE, KIF11 Y81F , and KIF11 ΔCterm neurons compared to NC-GFP. Kolmogorov-Smirnov Test. For graphs ( C , D ), error bars represent ± SEM. P-values are listed above respective comparisons. Source data are provided as a Source Data file.

Article Snippet: DIV16 primary hippocampal neurons culture (in 35 mm Mattek dishes) were transfected with 0.5 μg of NC-GFP (Origene TR30013) or KIF11-OE (Genecoepia CS-Mm03465-Lv236-01) for Supplementary Fig. ; or 0.5 μg NC-GFP (Origene TR30013), KIF11-OE (Genecoepia CS-Mm03465-Lv236-01), or Kif11 Y81F (Genecoepia CS-Mm03465-Lv236-02) for Supplementary Fig. . At DIV18, neuronal cultures were washed with PBS and then fixed in a freshly prepared solution of 4% paraformaldehyde for 15 min. After three or more rinses in PBS, the cells were then incubated in 10% normal horse serum(NHS) (Gibco) in PBS-T with 0.1% Triton X-100 for 1 h at room temperature to reduce the non-specific binding of the primary antibody.

Techniques: Expressing, Construct, Transfection

Journal: Nature Communications

Article Title: Intellectual disability-causing mutations in KIF11 impair microtubule dynamics and dendritic arborization

doi: 10.1038/s41467-026-70522-z

Figure Lengend Snippet: A Experimental timeline for in vivo expression of MCLID mutations in mouse. In utero electroporation (IUE) was used to insert eGFP, WT KIF11 (KIF11-OE), KIF11 Y81F , or KIF11 ΔCterm constructs into the hippocampi of embryonic day 15.5 mice. These mice developed until postnatal day 7 or day 21. Their brains were extracted, fixed in 4% PFA, cryoprotected, coronally sectioned at 50 µm, and imaged on a confocal microscope. B Representative images of neurons expressing eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm in the dorsal CA1 of P7 mice. C Sholl analysis of neurons expressing eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm in the dorsal CA1 of P7 mice. Two-way ANOVA followed by Tukey’s multiple comparisons test. D Number of branch points per neuron in P7 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. E Individual branch lengths per neuron in P7 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. F Sum of branch lengths per neuron in P7 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. N = 10(3),13(3),13(2),13(3) neurons (Mice) in eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm P7 mice, respectively. G Representative images of neurons expressing eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm in the dorsal CA1 of P21 mice. H Sholl analysis of neurons expressing eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm in the dorsal CA1 of P21 mice. Two-way ANOVA followed by Tukey’s multiple comparisons test. I Number of branch points per neuron in P21 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. J Individual branch lengths per neuron in P21 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. K Sum of branch lengths per neuron in P21 mice. One-way ANOVA followed by Tukey’s multiple comparisons test. N = 10(3),10(3),10(2), 10(3) neurons (Mice) in eGFP, KIF11-OE, KIF11 Y81F , or KIF11 ΔCterm P21 mice respectively. For all graphs ( D–F , I–K ), error bars represent ±SEM. P values are listed above respective comparisons. Source data are provided as a file.

Article Snippet: DIV16 primary hippocampal neurons culture (in 35 mm Mattek dishes) were transfected with 0.5 μg of NC-GFP (Origene TR30013) or KIF11-OE (Genecoepia CS-Mm03465-Lv236-01) for Supplementary Fig. ; or 0.5 μg NC-GFP (Origene TR30013), KIF11-OE (Genecoepia CS-Mm03465-Lv236-01), or Kif11 Y81F (Genecoepia CS-Mm03465-Lv236-02) for Supplementary Fig. . At DIV18, neuronal cultures were washed with PBS and then fixed in a freshly prepared solution of 4% paraformaldehyde for 15 min. After three or more rinses in PBS, the cells were then incubated in 10% normal horse serum(NHS) (Gibco) in PBS-T with 0.1% Triton X-100 for 1 h at room temperature to reduce the non-specific binding of the primary antibody.

Techniques: In Vivo, Expressing, In Utero, Electroporation, Construct, Microscopy