Journal: Frontiers in Immunology

Article Title: Rab7 inhibitor enhances stem cell differentiation into keratinocyte-like cells with anti-inflammatory properties

doi: 10.3389/fimmu.2025.1503007

Figure Lengend Snippet: Effect of Rab7 inhibition on the viability and morphology of ASCs. (A) Viability of ASCs after treatment with different concentrations of CID-1067700 at 24 and 72 hours. No significant changes in cell viability were observed. (B) Representative bright-field images illustrating the morphological changes in ASCs after 10 days of treatment with differentiation media (DM) with and without 40 μM CID-1067700, in comparison to undifferentiated ASCs. Scale bars = 100 μm.

Article Snippet: The adipose-derived mesenchymal stem cell (ASCs) line, ASC52telo (SCRC-4000, ATCC) immortalized with human telomerase reverse transcriptase (hTERT), were selected for their consistent stem cell properties and reproducibility.

Techniques: Inhibition, Comparison

Journal: Frontiers in Immunology

Article Title: Rab7 inhibitor enhances stem cell differentiation into keratinocyte-like cells with anti-inflammatory properties

doi: 10.3389/fimmu.2025.1503007

Figure Lengend Snippet: Rab7 inhibition leads to transcriptomic changes in ASCs. (A) Principal component analysis (PCA) of the transcriptomic profiles of cells treated as control (DM), with vehicle (DMSO), or with CID. PCA revealed clustering of the vehicle group with DM group, indicating a high degree of similarity between these two groups. (B) Volcano plot showed the differentially expressed genes (DEGs) in the CID-treated ASCs compared to those treated with DM. Downregulated genes are shown in blue, upregulated genes are in red, while insignificantly regulated genes are in grey.

Article Snippet: The adipose-derived mesenchymal stem cell (ASCs) line, ASC52telo (SCRC-4000, ATCC) immortalized with human telomerase reverse transcriptase (hTERT), were selected for their consistent stem cell properties and reproducibility.

Techniques: Inhibition, Control

Journal: Frontiers in Immunology

Article Title: Rab7 inhibitor enhances stem cell differentiation into keratinocyte-like cells with anti-inflammatory properties

doi: 10.3389/fimmu.2025.1503007

Figure Lengend Snippet: Impact of Rab7 inhibition on signalling pathways, biological processes, and upstream regulators in ASCs. (A) Chord Diagram of the top six significantly enriched pathways. (B) The top five enriched GO biological processes among the DEGs in the CID-treated ASCs. (C) Predicted activated or (D) inhibited upstream regulators for the DEGs in CID-treated ASCs compared to the control group. These predictions were made using the iPathwayGuide software.

Article Snippet: The adipose-derived mesenchymal stem cell (ASCs) line, ASC52telo (SCRC-4000, ATCC) immortalized with human telomerase reverse transcriptase (hTERT), were selected for their consistent stem cell properties and reproducibility.

Techniques: Inhibition, Control, Software

Journal: Frontiers in Immunology

Article Title: Rab7 inhibitor enhances stem cell differentiation into keratinocyte-like cells with anti-inflammatory properties

doi: 10.3389/fimmu.2025.1503007

Figure Lengend Snippet: Validation of the expression levels of the differentially expressed genes identified by microarray with qPCR. (A) Bar chart displaying the relative expression levels of seven selected DEGs ( COL81A, COL1A1, ITGB8, MGP, FGF7, VCAM1, and HMOX-1) . Bars represent the log2-fold changes in gene expression in CID-1067700-treated ASCs (CID) compared to the control group (DM). (B) Validation of the expression levels of the selected genes by qPCR. The expression levels are normalized to GAPDH and calculated relative to the expression of ASCs treated with DM only using the 2 -∆∆Cq method. Data are presented as mean ± standard deviation (SD) of three experimental replicates. The vehicle control was added to the qPCR to validate similarity with the negative control.

Article Snippet: The adipose-derived mesenchymal stem cell (ASCs) line, ASC52telo (SCRC-4000, ATCC) immortalized with human telomerase reverse transcriptase (hTERT), were selected for their consistent stem cell properties and reproducibility.

Techniques: Biomarker Discovery, Expressing, Microarray, Gene Expression, Control, Standard Deviation, Negative Control

Journal: Frontiers in Immunology

Article Title: Rab7 inhibitor enhances stem cell differentiation into keratinocyte-like cells with anti-inflammatory properties

doi: 10.3389/fimmu.2025.1503007

Figure Lengend Snippet: Rab7 Inhibition induces changes in the proteomic profile of ASCs. The upper left image shows the original scan of the protein array membrane. Noticeable reduction in the levels of inflammatory cytokines IL-1α, IL-8, IL-17A, and IL-32 were observed in CID-treated cells, whereas the expression level of EGF was increased. Bars represent the mean fold change in protein expression ± standard deviation (SD) of two experimental replicates in response to CID treatment compared to the control (DM).

Article Snippet: The adipose-derived mesenchymal stem cell (ASCs) line, ASC52telo (SCRC-4000, ATCC) immortalized with human telomerase reverse transcriptase (hTERT), were selected for their consistent stem cell properties and reproducibility.

Techniques: Inhibition, Protein Array, Membrane, Expressing, Standard Deviation, Control

Journal: Frontiers in Immunology

Article Title: Rab7 inhibitor enhances stem cell differentiation into keratinocyte-like cells with anti-inflammatory properties

doi: 10.3389/fimmu.2025.1503007

Figure Lengend Snippet: Rab7 Inhibition enhances the expression of epithelial differentiation markers in ASCs. The latter were cultured with differentiation media for ten days with and without CID-1067700, and immunocytochemistry analysis was performed to assess the expression of Cytokeratin 5 and 14, as well as Filaggrin and P63. Nuclei were counterstained with DAPI (blue). The bars indicate the mean fluorescence intensity unit (FU) per cell ± SD of 3 replicates. Asterisks represent significant difference compared to cells treated with differentiation media (DM) only (* p<0.5, ** p<0.01). Scale bars = 100 μm.

Article Snippet: The adipose-derived mesenchymal stem cell (ASCs) line, ASC52telo (SCRC-4000, ATCC) immortalized with human telomerase reverse transcriptase (hTERT), were selected for their consistent stem cell properties and reproducibility.

Techniques: Inhibition, Expressing, Cell Culture, Immunocytochemistry, Fluorescence

Journal: Frontiers in Immunology

Article Title: Rab7 inhibitor enhances stem cell differentiation into keratinocyte-like cells with anti-inflammatory properties

doi: 10.3389/fimmu.2025.1503007

Figure Lengend Snippet: Effect of Rab7 inhibition on the expression of keratinocyte markers in ASCs cells. The gene expression of (A) Vimentin, (B) Filaggrin, (C) Cytokeratin 5, and (D) Cytokeratin 14 was measured by Real-time PCR. The expression levels were normalized to GAPDH expression and calculated relative to cells treated with differentiation media (DM) alone. Asterisks indicate a significant difference compared to cells treated with DM (* p<0.05, ** p<0.01, *** p<0.001).

Article Snippet: The adipose-derived mesenchymal stem cell (ASCs) line, ASC52telo (SCRC-4000, ATCC) immortalized with human telomerase reverse transcriptase (hTERT), were selected for their consistent stem cell properties and reproducibility.

Techniques: Inhibition, Expressing, Gene Expression, Real-time Polymerase Chain Reaction

Journal: Journal of Personalized Medicine

Article Title: The Effect of Ionizing Irradiation on the Autotaxin-Lysophasphatidic Acid Axis and Interleukin-6/8 Secretion in Different Breast Cancer Cell Lines

doi: 10.3390/jpm14090968

Figure Lengend Snippet: Alterations in ATX expression on the mRNA level of ASC/TERT1 cells 24 h after stimulation with IL-6 ( A ) and IL-8 ( B ). The columns show the mean relative ATX expression (y-axis) in varying concentrations of IL-6 and IL-8 (x-axis) compared to YWHAZ and to the control group. Values were calculated using the 2 −ΔΔCT method, where 2 −ΔΔCT of control = 1. * p < 0.05 (Mann–Whitney U test).

Article Snippet: An ASC/TERT1 (human-adipose-tissue-derived telomerase-immortalized mesenchymal stem cell line) was purchased from Evercyte (Evercyte GmbH, Vienna, Austria) and cultivated in Endothelial Cell Growth Medium (EGM)-2 BulletKit (Lonza Group AG, Basel, Switzerland).

Techniques: Expressing, Control, MANN-WHITNEY

Journal: Biochemical pharmacology

Article Title: PPARγ antagonists induce aromatase transcription in adipose tissue cultures.

doi: 10.1016/j.bcp.2024.116095

Figure Lengend Snippet: Fig. 1. Lipid accumulation in human ASCs exposed to putative PPARγ antagonists during adipogenesis. Differentiation of human ASCs was induced with either high (2 μM) or low (0.1 μM) concentrations of rosiglitazone in the adipogenic medium. During differentiation, cells were treated with different environmental chemicals previously found to be PPARγ antagonists at concentrations of 0.25 to 5 μM or 0.5 to 10 μM. Lipids were stained with Oil Red O at day 12 of differentiation. Mi croscopy pictures show lipid accumulation in differentiated cells treated with different concentrations of putative PPARγ antagonists or solvent control (0 to 0.01 % DMSO). Lipid staining was quantified and presented in graphs showing study participant means ± SD (n = 4–6). Asterisks (*) indicate statistically significant dif ferences compared to differentiated solvent control cells at the same rosiglitazone concentration using Dunnett’s test (p < 0.05). (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The human A41 ASCs cell line (hTERT A41hWAT-SVF, passage 8–19) [21] and the mouse C3H10T1/2 mesenchymal stem cell line (CCL-226, ATCC, passage 6–11) were cultured in Dulbecco’s Modified Eagle Medium (DMEM; 41965–039, Gibco) containing 10 % FBS (F7524, SigmaAldrich) and 1 % PS solution (15070063, Gibco).

Techniques: Staining, Solvent, Control, Concentration Assay

Journal: Biochemical pharmacology

Article Title: PPARγ antagonists induce aromatase transcription in adipose tissue cultures.

doi: 10.1016/j.bcp.2024.116095

Figure Lengend Snippet: Fig. 2. Lipid droplet size and number in human ASCs exposed to putative PPARγ antagonists during adipogenesis. Differentiation of human ASCs was induced with adipogenic medium containing 2 μM rosiglitazone. During differentiation, cells were treated with previously identified PPARγ antagonists or solvent control (0 to 0.01 % DMSO). At day 12 of differentiation, lipids and nuclei were stained with LipidTOX and Hoechst, respectively. (A) Confocal microscopy pictures (study participant #8) show lipids in pink and nuclei in blue. Lipid droplet number and size was determined by image analysis of one field of view for each of the three study participants (#8, 10, 11). (B) Average lipid droplets per cell and (C) average lipid droplet diameter quantified by image analysis. Non-normalized data for all chemicals are shown for each study participant in B and C, where black is vehicle control and lighter shades of gray are higher concentrations of chemical. (D) Quantification of fluorescent stains using a plate reader and presented as LipidTOX normalized to Hoechst. All data are shown in graphs as study participant means ± SD (n = 3). Asterisks (*) indicate statistically significant differences compared to differentiated solvent control cells using Dunnett’s test (p < 0.05). (For inter pretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: The human A41 ASCs cell line (hTERT A41hWAT-SVF, passage 8–19) [21] and the mouse C3H10T1/2 mesenchymal stem cell line (CCL-226, ATCC, passage 6–11) were cultured in Dulbecco’s Modified Eagle Medium (DMEM; 41965–039, Gibco) containing 10 % FBS (F7524, SigmaAldrich) and 1 % PS solution (15070063, Gibco).

Techniques: Solvent, Control, Staining, Confocal Microscopy

Journal: Biochemical pharmacology

Article Title: PPARγ antagonists induce aromatase transcription in adipose tissue cultures.

doi: 10.1016/j.bcp.2024.116095

Figure Lengend Snippet: Fig. 3. Level of aromatase and adipocyte marker mRNA in response to inhibition of adipogenesis by PPARγ antagonists. Gene expression analysis of aromatase (CYP19A1) and adipocyte markers (PPARG, ADIPOQ, FASN) by RT-qPCR. (A) Human ASCs and adipocytes were directly isolated from human adipose tissue (n = 7). (B) A41 pre-adipocytes were undifferentiated or differentiated for 12 days (n = 8–10). (C) A41 cells at day 6 of differentiation were either returned to basal medium or kept in adipogenic medium for 2 days (n = 4). (D) A41 cells at day 6 (n = 3–4) or (E) at day 12 (n = 4) of differentiation were treated with 5 μM or 1 μM GW9662, respectively, during differentiation, and compared with a solvent control group (0.01 % DMSO or 0.002 %, respectively). (F) Mouse C3H10T1/2 cells were differentiated for 6 days in the presence or absence of 10 μM GW9662 or left undifferentiated. Gene expression analysis by RT-qPCR was performed for aromatase (Cyp19a1) using TaqMan assay and adipocyte markers (Pparg, Adipoq, Fabp4, and Slc2a4) using SYBR Green assay (n = 3). The graphs present means ± SD. Asterisk (*) and hash (#) indicate statistically significant differences compared to control cells using t test or two-way ANOVA, respectively (p < 0.05). Control cells are ASCs in (A), undifferentiated cells in (B), cells maintained in adipogenic medium in (C), and differentiated solvent control cells in (D-E). For C3H10T1/2 cells shown in (F), asterisk indicates statistically significant difference compared to differentiated control cells using two sample proportion test with Bonferroni correction (p < 0.05). ND: not detected.

Article Snippet: The human A41 ASCs cell line (hTERT A41hWAT-SVF, passage 8–19) [21] and the mouse C3H10T1/2 mesenchymal stem cell line (CCL-226, ATCC, passage 6–11) were cultured in Dulbecco’s Modified Eagle Medium (DMEM; 41965–039, Gibco) containing 10 % FBS (F7524, SigmaAldrich) and 1 % PS solution (15070063, Gibco).

Techniques: Marker, Inhibition, Gene Expression, Quantitative RT-PCR, Isolation, Solvent, Control, TaqMan Assay, SYBR Green Assay

Journal: Biochemical pharmacology

Article Title: PPARγ antagonists induce aromatase transcription in adipose tissue cultures.

doi: 10.1016/j.bcp.2024.116095

Figure Lengend Snippet: Fig. 6. Steroid hormone levels in response to PPARγ agonist or antagonist exposure. Hormone levels for a subset of steroids were measured in the culture medium of H295R cells using LC-MS/MS in response to treatment with 0.5 μM of GW9662 or zoxamide; 1 μM of forskolin, prochloraz, rosiglitazone, or pyridaben; 2 μM of pyraclostrobin; 5 μM of diphenyl phthalate or DEHPA; or solvent (0.005 % DMSO) for 48 h. (A) A heatmap shows the chemical-induced changes in the concentration of hormones secreted by H295R cells (n = 3). (B) The ratio of estrone to androstenedione and the ratio of estradiol to testosterone are used as measures of aromatase activity and are shown in bar plots. (C) Immunoblots show PPARγ protein levels in H295R cells compared to differentiated A41 adipocytes with vinculin as a reference protein. Data are presented as means ± SD. Asterisk (*) indicates statistically significant difference compared to solvent control cells using Dunnett’s test (p < 0.05).

Article Snippet: The human A41 ASCs cell line (hTERT A41hWAT-SVF, passage 8–19) [21] and the mouse C3H10T1/2 mesenchymal stem cell line (CCL-226, ATCC, passage 6–11) were cultured in Dulbecco’s Modified Eagle Medium (DMEM; 41965–039, Gibco) containing 10 % FBS (F7524, SigmaAldrich) and 1 % PS solution (15070063, Gibco).

Techniques: Liquid Chromatography with Mass Spectroscopy, Solvent, Concentration Assay, Activity Assay, Western Blot, Control