Journal: Cell Death & Disease

Article Title: Targeting ENO1 reprograms macrophage polarization to trigger antitumor immunity and improves the therapeutic effect of radiotherapy

doi: 10.1038/s41419-026-08416-7

Figure Lengend Snippet: A The expression of total and surface ENO1 in human CRC cell lines was evaluated by immunoblotting and flow cytometry. B The expression of total and surface ENO1 in human BRCA cell lines was evaluated by immunoblotting and flow cytometry. C HT29 and LoVo cells were treated with different doses of recombinant human TGFβ1 protein (rhTGFβ1) for 24 hr, after which surface ENO1 expression was analyzed via flow cytometry. D HS578T and MDA-MB-468 cells were treated with different doses of the rhTGFβ1 protein for 24 hr, after which the surface ENO1 level was analyzed via flow cytometry. E LoVo cells were treated with LPS (1 μg/mL), rhTGFβ1 protein (10 ng/mL) or H 2 O 2 (50 μM) in combination with a PRMT5 inhibitor (AMG-193, 1 μM) or a PRMT6 inhibitor (EPZ020411 hydrochloride, 1 μM) for 24 hr. The surface ENO1 level was analyzed via flow cytometry. * p < 0.05, ** p < 0.01 and *** p < 0.001. One-Way ANOVA test ( n = 3). F HT29 cells were treated with rhTGFβ1 protein in combination with a PRMT5 inhibitor (1 μM) for 24 hr. The surface ENO1 expression was analyzed via flow cytometry. ** p < 0.01 and *** p < 0.001. One-Way ANOVA test ( n = 3). G LoVo shNC and LoVo shPRMT5 cells were treated with LPS (1 μg/mL), rhTGFβ1 protein (10 ng/mL) or H 2 O 2 (50 μM) for 24 hr. The surface level of ENO1 was examined by flow cytometry. * p < 0.05, ** p < 0.01 and *** p < 0.001. One-Way ANOVA test ( n = 3). H HT29 and LoVo cells were treated with LPS (1 μg/mL) and RT (5 Gy) for 24 hr. The surface level of ENO1 was examined by flow cytometry. * p < 0.05 and ** p < 0.01. One-Way ANOVA test ( n = 3). I HT29 cells were treated with RT (5 Gy) in combination with a TGFβR1 inhibitor (1 μM), a Smad3 inhibitor (1 μM), or a PRMT5 inhibitor (1 μM) for 24 hr. The surface level of ENO1 was examined by flow cytometry. *** p < 0.001. One-Way ANOVA test ( n = 3). J HT29 cells were treated with RT (5 Gy) in combination with a TGFβR1 inhibitor (1 μM), a Smad3 inhibitor (1 μM), or a PRMT5 inhibitor (1 μM) for 24 hr. The level of plasmin activity was examined via an ELISA kit. * p < 0.05. One-Way ANOVA test ( n = 3). K MDA-MB-468 cells were treated with RT (5 Gy) in combination with a TGFβR1 inhibitor (1 μM), a Smad3 inhibitor (1 μM), or a PRMT5 inhibitor (1 μM) for 24 hr. The surface level of ENO1 was examined by flow cytometry. * p < 0.05, ** p < 0.01 and *** p < 0.001. One-Way ANOVA test ( n = 3). L MDA-MB-468 cells were treated with RT (5 Gy) in combination with a TGFβR1 inhibitor (1 μM), a Smad3 inhibitor (1 μM), or a PRMT5 inhibitor (1 μM) for 24 hr. The level of plasmin activity was examined via an ELISA kit. * p < 0.05, and ** p < 0.01. One-Way ANOVA test ( n = 3). M HT29 cells were treated with RT (5 Gy) in combination with a PRMT5 inhibitor (1 μM) for 24 hr. The level of surface-methylated ENO1 was evaluated by immunoprecipitation and immunoblotting.

Article Snippet: The TGFβR1 inhibitor galunisertib (HY-13226, MCE, USA) [ ], Smad3 inhibitor SIS3 (HY-13013, MCE, USA) and PRMT5 inhibitor EPZ015666 (HY-12727, MCE, USA) were dissolved in DMSO to a concentration of 10 mM.

Techniques: Expressing, Western Blot, Flow Cytometry, Recombinant, Activity Assay, Enzyme-linked Immunosorbent Assay, Methylation, Immunoprecipitation

Journal: Cell Death & Disease

Article Title: Targeting ENO1 reprograms macrophage polarization to trigger antitumor immunity and improves the therapeutic effect of radiotherapy

doi: 10.1038/s41419-026-08416-7

Figure Lengend Snippet: A A total of 5 × 10 5 CT26 cells were subcutaneously injected into the left legs of BALB/c mice for 5 days and then intraperitoneally administered with anti-ENO1 antibodies (HuL001, 40 mg/kg) or clodronate liposomes (50 μL/mouse) on the indicated days ( n = 5). Local radiotherapy was given on Day 10. The tumor volume was recorded every three days. * p < 0.05. Two-Way ANOVA test ( n = 4). B The densities of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors were analyzed by immunofluorescence staining. The quantification of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors. * p < 0.05 and *** p < 0.001. One-Way ANOVA test ( n = 3). C 4T1 cells (5 ×10 4 ) were subcutaneously injected into the left legs of BALB/c mice for 4 days and then intraperitoneally administered anti-ENO1 antibodies (HuL001, 20 mg/kg) six times on the indicated days ( n = 5). Local radiotherapy was given on Days 10 and 12. The tumor volume was recorded every three days. * p < 0.05 and *** p < 0.001. Two-Way ANOVA test ( n = 4). D The resected tumors were weighed on Day 40. * p < 0.05. One-Way ANOVA test ( n = 4). E The densities of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors were analyzed by immunofluorescence staining. F The quantification of M1 (CD80 + ) and M2 (CD206 + ) macrophages in resected tumors. * p < 0.05. One-Way ANOVA test ( n = 3). G The frequencies of M1 (CD11c + CD11b + F4/80 + CD45 + 7AAD - CD3 - CD19 - ) and M2 (CD1206 + CD11b + F4/80 + CD45 + 7AAD - CD3 - CD19 - ) tumor-infiltrating macrophages were analyzed by flow cytometry. * p < 0.05. One-Way ANOVA test ( n = 3-4). H The quantification of the M1/M2 ratio is shown. * p < 0.05. One-Way ANOVA test ( n = 3). I A representative image of flow cytometric analysis of GzmB + (GzmB hi CD8 + CD3 + CD45 + 7AAD - ) T cells. J The frequency of GzmB + (GzmB hi CD8 + CD3 + CD45 + 7AAD - ) T cells was analyzed by flow cytometry. * p < 0.05. One-Way ANOVA test ( n = 3-4). K The density of GzmB + (GzmB hi CD8 + CD3 + CD45 + 7AAD - ) T cells is shown. * p < 0.05. One-Way ANOVA test ( n = 3-4). L The density of IFNγ + (IFNγ hi CD8 + CD3 + CD45 + 7AAD - ) T cells is shown ( n = 3-4). * p < 0.05 and ** p < 0.01. One-Way ANOVA test ( n = 3-4). M The proposed mechanism of TGFβ1/TGFβR/Smad3/PRMT5-mediated ENO1 translocation for lactate release via MCT4.

Article Snippet: The TGFβR1 inhibitor galunisertib (HY-13226, MCE, USA) [ ], Smad3 inhibitor SIS3 (HY-13013, MCE, USA) and PRMT5 inhibitor EPZ015666 (HY-12727, MCE, USA) were dissolved in DMSO to a concentration of 10 mM.

Techniques: Injection, Liposomes, Immunofluorescence, Staining, Flow Cytometry, Translocation Assay

Journal: bioRxiv

Article Title: PRMT5 inhibitors actively promote metastatic progression of lung adenocarcinoma

doi: 10.64898/2026.01.30.702866

Figure Lengend Snippet: a. Schematic of LUAD dedifferentiation spectrum, including the relative states of KP Early and KP Late . Each bar represents the general expression pattern of the given dedifferentiation state marker. b. Schematic of generation of PRMT5i-resistant variants of KP Late (KP1 36 ) and KP Early (KP2 36 ) cell lines. c. Dose response curves for KP Late (blue) and a representative KP Late -R line, KP Late -R1 (red) treated with JNJ-64619178 (PRMT5i) for 5 days. Data are mean ± SD of 3 technical replicates/line. p=0.0012, Welch’s t-test. d. GSEA results for gene sets listed in Supplementary Fig. 1c comparing KP Early and KP Late to their resistant lines. Color represents normalized enrichment score, while size inversely correlates with significance. e. Western blot of core LUAD dedifferentiation markers (Nkx2.1, Hnf4a, Hmga2) and Hsp90 loading control in KP Early and KP Late , as well as one control (DMSO selection) and three PRMT5i-resistant populations. f. UMAP of scRNA-seq dataset from an autochthonous KP model of LUAD with ( Left panel ) all clusters marked, ( Center panel ) Stmn2 expression, or ( Right panel ) Sox11 expression within the individual clusters. Cluster 12 (circled) was previously described to contain late-stage, pro-metastatic cells 44 .

Article Snippet: The PRMT5 inhibitor JNJ-64619178 was used for all PRMT5i treatment experiments and obtained from MedChemExpress (HY-101564).

Techniques: Expressing, Marker, Western Blot, Control, Selection

Journal: bioRxiv

Article Title: PRMT5 inhibitors actively promote metastatic progression of lung adenocarcinoma

doi: 10.64898/2026.01.30.702866

Figure Lengend Snippet: a. Differentially accessible loci in KP Early after 5-day vehicle or 100nM PRMT5i treatment. Points represent individual loci that are more accessible (pink, p adj <0.01, log 2 FC>1), less accessible (blue, p adj <0.01, log 2 FC<-1), or not differentially accessible (gray). b. Cumulative fraction of the coefficient of variation for significant loci of KP Early after 5-day vehicle or PRMT5i treatment, indicating vehicle-derepressed (blue) and PRMT5i-derepressed (red) loci. p<10 -44 , F test. c. ChromVAR deviation score differences between vehicle- and PRMT5i-treated parental cells (x-axis) and resistant and parental cells (y-axis) for KP Early and KP Late . Points indicate differentially accessible (dark gray or colored, p adj <0.01) and not significant (light gray) motifs. Colored points represent motifs within the same TF family. d. Accessibility of indicated TF family motifs across: ( Top panel ) UMAP projections of a scATAC-seq dataset from an autochthonous model of LUAD, and ( Bottom panel ) chromVAR deviation scores for each biological replicate (n=3) of parental (blue), PRMT5i-treated parental (pink), and resistant (red) lines. e. Four categories of genome-wide representation of loci indicating derepression or repression of peak accessibility in response to a 5-day PRMT5i treatment of KP Early and whether this is established or not, in the stably-resistant KP Early -R1. ( Left panel ) Representative accessibility track, ( Center panel ) genome-wide accessibility heatmap, and ( Right panel ) all enriched TF motifs (p<10 -50 ) are shown. Colors denote: parental (blue), PRMT5i-treated parental (pink), and stable resistant (red) lines. f. Comparison of loci that show differentially accessible in response to a 5-day PRMT5i treatment of KP Early (drug-responsive) versus ones established in stably-resistant cells for ( Top panel ) KP Early and ( Bottom panel ) KP Late . Proportion of drug-responsive peaks that are also resistance state peaks is shown as a percentage. g. CTCF chromVAR deviation score in KP Early and KP Late treated with vehicle (blue) or PRMT5i for 5 days (pink). h. Genomic annotation of PRMT5-derepressed loci in KP Early and KP Late .

Article Snippet: The PRMT5 inhibitor JNJ-64619178 was used for all PRMT5i treatment experiments and obtained from MedChemExpress (HY-101564).

Techniques: Genome Wide, Stable Transfection, Comparison

Journal: bioRxiv

Article Title: PRMT5 inhibitors actively promote metastatic progression of lung adenocarcinoma

doi: 10.64898/2026.01.30.702866

Figure Lengend Snippet: a. Schematic of the CDKN2A and adjacent MTAP loci indicating gene products and functions. b. Frequents of CDKN2A copy number losses in various tumor types, with SKCM (skin cutaneous melanoma) and LUAD highlighted. c. Schematic of effects of CDKN2A- deficiency on MTAP and PRMT5 biology. d. Normalized RNA expression counts for HMGA2 and RUNX2 in CDKN2A / MTAP -deficient LU99 cells treated with vehicle (blue) or MRTX1719 (red), an MTA-cooperative PRMT5i, for 3 or 5 days. Data are mean ± SD of 2 biological replicates/line. *p<0.05, **p<0.01, ***p<0.001, Student’s t-test. e-f. Differentially expressed genes comparing CDKN2A DKO to the rest of the tumors (+) in the TCGA LUAD ( e ) Firehose Legacy or ( f ) PanCancer cohorts. ( Left panel ) Points represent individual genes that have increased expression (red, p<0.05, log 2 FC>0.5) or decreased expression (blue, p<0.05, log 2 FC<-0.5). ( Right panel ) Expression of HMGA2 in indicated groups. Points represent individual patients with the total number of patients per group listed below the axis. p=0.00068 Legacy cohort and p=0.00003 PanCancer cohort, Student’s t-test.

Article Snippet: The PRMT5 inhibitor JNJ-64619178 was used for all PRMT5i treatment experiments and obtained from MedChemExpress (HY-101564).

Techniques: RNA Expression, Expressing