Journal: iScience

Article Title: Dissecting genotype-specific effects of disease-associated genetic variants

doi: 10.1016/j.isci.2026.116143

Figure Lengend Snippet: Overview of experimental workflow Affinity purification MS were completed on the undifferentiated KOLF2.1J WT cell line. Bulk RNA-seq and ONT long-read WGS + 5 mC methylation analysis was completed on undifferentiated KOLF2.1J clones containing the following rs11610045 genotypes: A|A (WT), G|G (edited), and A|A (reversed). WT and edited (but not reverse edited) clones were differentiated to iPSC-derived cortical neurons and bulk RNA-seq completed. iPSC, induced pluripotent stem cell; sgRNA, single-guide ribonucleic acid; ssODN, single-strand oligodeoxynucleotide; RNP, ribonucleoprotein; HDR enhancer, homology-directed repair enhancer; ONT, Oxford Nanopore Technologies; WGS, whole-genome sequencing.

Article Snippet: The KOLF2.1J iPSC line , was obtained from Jackson laboratory ( https://www.jax.org/jax-mice-and-services/ipsc ).

Techniques: Affinity Purification, RNA Sequencing, Methylation, Clone Assay, Derivative Assay, Sequencing

Journal: iScience

Article Title: Dissecting genotype-specific effects of disease-associated genetic variants

doi: 10.1016/j.isci.2026.116143

Figure Lengend Snippet: Restoring the rs11610045 G|G genotype to A|A also restores gene expression for a subset of genes (A) CRISPR-Cas9 editing of the rs11610045 G|G genotype to A|A restored the expression of 75 genes to WT levels. Green bars represent the log2FC when comparing the edited rs11610045 G|G genotype with that of the A|A genotype (WT) KOLF2.1J cells. Blue bars represent the log2FC when comparing gene expression in the rs11610045 A|A (reversed) with that of the rs11610045 G|G genotype (edit) clones. In all instances, log2FC > 1 and Bonferroni adj. p value <0.05. (B) Expression of CEL , PDGFB , and THBS2 are rs11610045 genotype dependent and have been linked to PD. Boxplots show the median (center line), interquartile range (box), and whiskers extending to 1.5× the interquartile range. Individual data points are overlaid. (C) PDGFB interacts with THBS1 in a protein-protein interaction network (StringDB, high confidence interaction score ≥0.700).

Article Snippet: The KOLF2.1J iPSC line , was obtained from Jackson laboratory ( https://www.jax.org/jax-mice-and-services/ipsc ).

Techniques: Gene Expression, CRISPR, Expressing, Clone Assay

Journal: Frontiers in Immunology

Article Title: STAT6 inhibition of M2 macrophages suppresses tumor growth by modulating the tumor microenvironment in colon cancer model

doi: 10.3389/fimmu.2026.1733991

Figure Lengend Snippet: TAMpep-IP reduces phosphorylation of STAT6 in M2 macrophages. (A) Schematic structure of TAMpep-IP composed of an M2 macrophage-homing peptide (TAMpep), a cleavable linker, and a STAT6-inhibitory peptide (IP). (B) THP-1 cells were differentiated into M0, M1, or M2 macrophages and stained for phosphorylated STAT6 (p-STAT6; red). Nuclear staining was performed with DAPI (blue). Immunofluorescence microscopy revealed elevated nuclear p-STAT6 in M2 macrophages compared to M0 and M1. Representative confocal images were acquired using a 40× objective lens. Scale bars, 20 μm. (C) M0 and M2 macrophages were treated with TAMpep, IP, or TAMpep-IP (0.5 μM, 72 h). Flow cytometry was used to measure p-STAT6 expression levels (mean fluorescence intensity), showing that TAMpep-IP significantly reduced p-STAT6 in M2 macrophages. (D) Western blot analysis was performed to detect p-STAT6 and total STAT6 levels in M2 macrophages after administration with TAMpep, IP, or TAMpep-IP (0.5 μM, 72 h). TAMpep-IP effectively reduced p-STAT6. The experiment was performed in triplicate. All data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p < 0.001 and ****p < 0.0001..

Article Snippet: The human monocytic leukemia cell line THP-1 (KCLB; Korean Cell Line Bank, Seoul, Korea) was cultured in RPMI 1640 medium (Welgene, Gyeongsangbuk, Korea) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% penicillin–streptomycin (Hyclone, Logan, UT, USA).

Techniques: Phospho-proteomics, Staining, Immunofluorescence, Microscopy, Flow Cytometry, Expressing, Fluorescence, Western Blot

Journal: Frontiers in Immunology

Article Title: STAT6 inhibition of M2 macrophages suppresses tumor growth by modulating the tumor microenvironment in colon cancer model

doi: 10.3389/fimmu.2026.1733991

Figure Lengend Snippet: TAMpep-IP inhibits polarization of M2 macrophages. (A) THP-1 monocytes were differentiated into M0 and M2 macrophages, and M2 cells were treated with TAMpep-IP (0.5 μM, 72 h). Quantitative RT-PCR analysis showed that TAMpep-IP significantly reduced the mRNA expression of TGF-β and Arg-1, two markers associated with M2 polarization. (B) ELISA was performed to measure the levels of secreted TGF-β and IL-13 in the culture supernatant of M0, M2, and TAMpep-IP–treated M2 macrophages (72 h). TAMpep-IP markedly decreased secretion of both cytokines. (C) Flow cytometric analysis was used to assess CD206 surface expression in M0, M2, and TAMpep-IP–treated M2 macrophages (72 h). CD206 expression was significantly decreased in the TAMpep-IP group compared to untreated M2 macrophages. All data are presented as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p < 0.0001. (D) The expression of IL-1β mRNA was analyzed by quantitative RT-PCR in M0, M1, and M2 macrophages after TAMpep-IP. TAMpep-IP significantly upregulated IL-1β expression in M2 macrophages, to levels comparable with M1-polarized cells.

Article Snippet: The human monocytic leukemia cell line THP-1 (KCLB; Korean Cell Line Bank, Seoul, Korea) was cultured in RPMI 1640 medium (Welgene, Gyeongsangbuk, Korea) supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% penicillin–streptomycin (Hyclone, Logan, UT, USA).

Techniques: Quantitative RT-PCR, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Regenerative Therapy

Article Title: WTAP stabilizes MMP12 expression to promote the malignant phenotypes of esophageal cancer cells

doi: 10.1016/j.reth.2026.101101

Figure Lengend Snippet: MMP12 silencing inhibited M2 macrophage polarization. THP-1 cells were differentiated into M0 macrophages (THP-1 M0) by treatment with 100 ng/mL PMA for 24 h. (A) Flow cytometry was used to quantify the number of CD68-positive cells. Subsequently, KYSE150 cells were co-cultured with the THP-1-derived macrophages using a Transwell system. (B) The mRNA levels of IL-10, Arg-1, and TGF-β were detected by qRT-PCR. (C) Flow cytometry was used to quantify the number of CD206-positive macrophages. (D) Cell migration analysis by transwell migration assay. ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: For immunophenotyping, single-cell suspensions of both THP-1 and THP-1-M0 cells were labeled with an anti-CD68 antibody (E-AB-F1299L, Elabscience, Wuhan, China).

Techniques: Flow Cytometry, Cell Culture, Derivative Assay, Quantitative RT-PCR, Migration, Transwell Migration Assay

Journal: Regenerative Therapy

Article Title: WTAP stabilizes MMP12 expression to promote the malignant phenotypes of esophageal cancer cells

doi: 10.1016/j.reth.2026.101101

Figure Lengend Snippet: WTAP silencing inhibited M2 macrophage polarization by regulating MMP12. THP-1 cells were differentiated into M0 macrophages (THP-1 M0) by treatment with 100 ng/mL PMA for 24 h. KYSE150 cells were transfected with si-WTAP, MMP12 overexpression plasmid, or the matched control (si-NC and oe-NC). Subsequently, these KYSE150 cells were co-cultured with the THP-1-derived macrophages using a Transwell system. (A) The mRNA levels of IL-10, Arg-1, and TGF-β were detected by qRT-PCR. (B) Flow cytometry was used to quantify the number of CD206-positive macrophages. (C) Cell migration analysis by transwell migration assay. ∗ P < 0.05, ∗∗ P < 0.01 and ∗∗∗ P < 0.001.

Article Snippet: For immunophenotyping, single-cell suspensions of both THP-1 and THP-1-M0 cells were labeled with an anti-CD68 antibody (E-AB-F1299L, Elabscience, Wuhan, China).

Techniques: Transfection, Over Expression, Plasmid Preparation, Control, Cell Culture, Derivative Assay, Quantitative RT-PCR, Flow Cytometry, Migration, Transwell Migration Assay