Journal: JACC: Basic to Translational Science

Article Title: Small Interfering RNA Therapy Targeting the Long Noncoding RNA SMILR for Therapeutic Intervention in Coronary Artery Bypass Graft Failure

doi: 10.1016/j.jacbts.2025.101364

Figure Lengend Snippet: Transcriptomic Profiling of BHF7-Transfected Smooth Muscle Cells Demonstrates that BHF7 Targets a Core Cell Cycle Gene Network (A) Principle component analysis of saphenous vein smooth muscle cells transfected with lipofectamine RNAiMax alone (mock), siNTC, or BHF7 to a final dose of 25 nmol/L. (B) BHF7 targets a cell cycle-enriched gene network. Our previous research identified a “SMILR-dependent network.” The top 20 differentially-expressed genes identified from this network are enriched for cell cycle-associated genes. This same set of SMILR-dependent genes demonstrate significantly reduced expression when BHF7 is transfected into cells. (C) RNA-sequencing expression analysis of CENPF from NT, mock-transfected, IL1-PDGF-treated cells (mock), siNTC, or cells transfected with BHF7 to a final dose of 25 nmol/L. (D) qRT-PCR validation of RNA-sequencing data for CENPF expression after transfection as indicated. Each colored dot represents smooth muscle cells derived from a single patient. N = 4, ∗ P < 0.05, ∗∗ P < 0.01, repeated measures analysis of variance w/Dunnett’s test for multiple comparisons. (E) Gene ontology (GO), reactome, and KEGG pathway analysis of BHF7-repressed genes reveals a broad network of genes associated with cell cycle progression and mitosis. Abbreviations as in and .

Article Snippet: Six hours post-transfection, cells were quiesced for 48 hours in Dulbecco’s Modified Eagle’s Medium (DMEM) (Gibco, Thermo Fisher Scientific) supplemented with 50 μg/mL penicillin (Invitrogen, Thermo Fisher Scientific), 50 μg/mL streptomycin (Invitrogen, Thermo Fisher Scientific) and 0.2% [v/v] fetal bovine serum (FBS) (Gibco, Thermo Fisher Scientific) and then stimulated for a further 48 hours with fresh 0.2% FBS DMEM media containing 10 ng/mL IL1α (R&D Systems 200-LA-010) and 20 ng/mL PDGF-ββ (R&D Systems 220-BB-010), and 10 μmol/L 5-ethynyl-2-deoxyuridine (EdU) (Invitrogen A10044, Thermo Fisher Scientific) where proliferation was assessed.

Techniques: Transfection, Expressing, RNA Sequencing, Quantitative RT-PCR, Biomarker Discovery, Derivative Assay

Journal: Biomolecules

Article Title: The Contribution of CD26-Negative Fibroblasts to Endometrial Scarring

doi: 10.3390/biom15101433

Figure Lengend Snippet: Effect of IL1α on CD26 expression in primary endometrial stromal cells. Cells were stimulated with increasing concentrations of IL1α (5–50 ng/mL) for 24 h ( A ) and 48 h ( B ) and CD26 protein levels analyzed by Western blot. Actin was used as loading control. IL1α significantly increased CD26 expression after 48 h at 20 ng/mL and 50 ng/mL. Representative immunoblots and the corresponding densitometric quantification from 3 independent experiments are shown. ( C ) Effect of IL1α and the CD26 inhibitor DPA on CD26 protein levels in stromal cells. Cells were treated with increasing concentrations of DPA and IL1α for 48 h. CD26 protein levels were analyzed by Western blot. IL1α significantly induced CD26, whereas DPA could not abrogate its effects. Unstimulated cells were set to 100% and used as a control. Each bar represents the mean ± SEM of three independent experiments performed in duplicates. ** p < 0.01; *** p < 0.001; Ctrl, control; DPA, diprotin A; n.s, not significant. Original images can be found in .

Article Snippet: The following materials were used: recombinant human IL1α (cat-no. 200-LA/CF; R&D Systems); anti-actin (cat-no. A3853, Sigma Aldrich), anti-mouse conjugated secondary antibody (cat-no. 7067) and anti-rabbit conjugated secondary antibody (cat-no. 7074) were all from Cell signaling technology (Frankfurt, Germany).

Techniques: Expressing, Western Blot, Control

Journal: Biomolecules

Article Title: The Contribution of CD26-Negative Fibroblasts to Endometrial Scarring

doi: 10.3390/biom15101433

Figure Lengend Snippet: Effects of IL1α and DPA on wound healing in vitro of primary endometrial stromal cells. ( A ) The confluent monolayer of stromal cells was disrupted by scratching with a pipette tip ( upper panel , 0 h) and incubated with DPA and IL1α. Cell-free areas were monitored 24 h after DPA and IL1α treatments by capturing images ( lower panel , 24 h). ( B ) The results are presented as percentages of the free gaps quantified with ImageJ. IL1α promoted wound healing of the stromal cells in vitro, which was inhibited by DPA treatment. Values are presented as means ± SEM of three independent experiments performed in duplicates. ** p < 0.01; Ctrl, control: DPA, diprotin A.

Article Snippet: The following materials were used: recombinant human IL1α (cat-no. 200-LA/CF; R&D Systems); anti-actin (cat-no. A3853, Sigma Aldrich), anti-mouse conjugated secondary antibody (cat-no. 7067) and anti-rabbit conjugated secondary antibody (cat-no. 7074) were all from Cell signaling technology (Frankfurt, Germany).

Techniques: In Vitro, Transferring, Incubation, Control

Journal: Biomolecules

Article Title: The Contribution of CD26-Negative Fibroblasts to Endometrial Scarring

doi: 10.3390/biom15101433

Figure Lengend Snippet: Effects of IL1α and DPA on secretion of COL1A1 and TGF-β3 of primary endometrial stromal cells. Cells were incubated with DPA and IL1α for 48 h. Secretion of COL1A1 and TGF-β3 was quantified by ELISAs. ( A ) IL1α dose-dependently increased COL1A1 secretion, which was significantly abrogated by DPA. ( B ) IL1α decreased secretion of TGF-β3, which was significantly attenuated by DPA. Untreated cells were set to 100% and used as controls. Each bar represents the means ± SEM of three independent experiments performed in duplicates. * p ≤ 0.05; ** p < 0.01; Ctrl, control; DPA, diprotin A; COL1A1, pro-collagen 1A1.

Article Snippet: The following materials were used: recombinant human IL1α (cat-no. 200-LA/CF; R&D Systems); anti-actin (cat-no. A3853, Sigma Aldrich), anti-mouse conjugated secondary antibody (cat-no. 7067) and anti-rabbit conjugated secondary antibody (cat-no. 7074) were all from Cell signaling technology (Frankfurt, Germany).

Techniques: Incubation, Control

Journal: Biomolecules

Article Title: The Contribution of CD26-Negative Fibroblasts to Endometrial Scarring

doi: 10.3390/biom15101433

Figure Lengend Snippet: Effects of IL1α and DPA on migration and viability of primary endometrial stromal cells. ( A ) Stromal cells were incubated with DPA and IL1α for 24 h. IL1α enhanced endometrial stromal cell migration, which was inhibited by DPA. Untreated cells were set to 100% and used as a control (Ctrl). ( B ) Stromal cells were incubated with DPA and IL1α for 48 h. Cell viability was assessed by the trypan blue exclusion assay. DPA suppressed IL1α-induced cell viability. Untreated cells were set to 100% and used as a control. The values are presented as means ± SEM of three independent experiments performed in duplicates. Ctrl, control; DPA, diprotin A. * p ≤ 0.05; ** p < 0.01.

Article Snippet: The following materials were used: recombinant human IL1α (cat-no. 200-LA/CF; R&D Systems); anti-actin (cat-no. A3853, Sigma Aldrich), anti-mouse conjugated secondary antibody (cat-no. 7067) and anti-rabbit conjugated secondary antibody (cat-no. 7074) were all from Cell signaling technology (Frankfurt, Germany).

Techniques: Migration, Incubation, Control, Trypan Blue Exclusion Assay

Journal: International Journal of Cancer

Article Title: ARP2 /3 complex affects myofibroblast differentiation and migration in pancreatic ductal adenocarcinoma

doi: 10.1002/ijc.35246

Figure Lengend Snippet: ARP2/3 complex is coupled with myofibroblast differentiation. (A), (B) This UMAP plot visually represents the presence of two distinct fibroblast populations (iCAFs and myCAFs) by using the published markers. (C) The dot plot illustrates the differential expression of the Arp2/3 complex signature in iCAFs and myCAFs, shedding light on their distinct molecular characteristics (lower left). (D) Immunofluorescence co‐staining of PDAC tumor tissue with ARP3 (green), αSMA (red) and DAPI (blue). One representative image is shown ( n = 8). Scale bars: 100 μm. (E) Correlation analysis of αSMA+ and ARP3+ cells was conducted using the “Colocalization Finder” tool in ImageJ 1.5.3. (F) This model provides insight into the signaling pathways that influence the formation of iCAFs and myCAFs in PDAC: TGFβ signaling plays a key role in myCAF formation, IL1 signaling drives iCAF development. (G) The results from qRT‐PCR analysis reveal the expression levels of myCAF markers (ACTA2, TAGLN, and MYL9) when treated with TGFβ1 (20 ng/mL) and iCAF markers (CXCL1, IL6, and CCL2) when treated with IL1α (1 ng/mL) in human PSCs, both in the presence and absence of CK666. These findings are derived from three independent experiments and are presented as mean ± SEM. *, indicates statistical significance with p <.05, as determined by a Student t ‐test.

Article Snippet: Commercially available reagents were used, including CK666 (#SML00006, Sigma‐Aldrich, Munich, Germany), human TGFβ1 (#T7039‐2UG, Sigma‐Aldrich), and human IL1α (#200‐LA‐002/CF, R&D Systems, Minneapolis, USA).

Techniques: Quantitative Proteomics, Immunofluorescence, Staining, Protein-Protein interactions, Quantitative RT-PCR, Expressing, Derivative Assay