Journal: Cell Cycle

Article Title: M2 macrophages promote PKM2 production in fibroblasts to alleviate UVB-induced photoaging

doi: 10.1080/15384101.2025.2514988

Figure Lengend Snippet: CCL1 secreted by M2 macrophages enhances PKM2 production in photoaged L929 cells. (a) Comparison of CCL1 levels in M1/M2 macrophages and their supernatants. (b) Immunofluorescence staining shows that M2 macrophages produce higher levels of CCL1. (c,d) Western blot and immunofluorescence staining reveal that both L929 and photoaged L929 cells express CCR8. (e) Exogenous CCL1 reduces the proportion of SA-β-GAL positive senescent cells. (f, g) Exogenous CCL1 increases the production of collagen and PKM2, while reducing the activation of P16, P21, P53, Caspase-3, and ATM in photoaged L929 cells. (h, i) The CCR8 antagonist R243 partially reverses the effects of M2 macrophage CM on photoaged L929 cells. If variances were equal, comparisons between two groups were conducted using the unpaired t-test, and comparisons among multiple groups were performed using one-way ANOVA. If variances were unequal, Welch’s correction was applied to reanalyze the data. For data that did not meet the assumption of normality, the Mann–Whitney U-test was used for two-group comparisons, and the Kruskal–Wallis test was applied for comparisons among multiple groups. n ≥ 3 (biological replicates).

Article Snippet: When cells reached 50%–60% confluence, the medium was replaced with fresh serum-free medium, and cells were treated for 48 h with various components, including macrophage-conditioned media (CM), PKM2-IN-1 (MedChemExpress, USA), CCL1 (MedChemExpress, USA), and R243 (MedChemExpress, USA).

Techniques: Comparison, Immunofluorescence, Staining, Western Blot, Activation Assay, MANN-WHITNEY

Journal: Cell Cycle

Article Title: M2 macrophages promote PKM2 production in fibroblasts to alleviate UVB-induced photoaging

doi: 10.1080/15384101.2025.2514988

Figure Lengend Snippet: M2 macrophages activate the TGF-β1/SMAD2 pathway in L929 cells to alleviate photoaging. (a) Activation of the TGF-β1/SMAD2 pathway is reduced in photoaged L929 cells. (b, c) PKM2 inhibitor PKM2-IN-1 blocks the activation of the TGF-β1/SMAD2 pathway in L929 cells, whereas the overexpression of PKM2 enhances this activation. (d, e, f) Addition of M2 macrophage CM or exogenous CCL1 promotes activation of the TGF-β1/SMAD2 pathway in photoaged L929 cells, while the CCR8 antagonist R243 inhibits the effect of M2 macrophage CM. If variances were equal, comparisons between two groups were conducted using the unpaired t-test, and comparisons among multiple groups were performed using one-way ANOVA. If variances were unequal, Welch’s correction was applied to reanalyze the data. For data that did not meet the assumption of normality, the Mann–Whitney U-test was used for two-group comparisons, and the Kruskal–Wallis test was applied for comparisons among multiple groups. n≥3 (biological replicates).

Article Snippet: When cells reached 50%–60% confluence, the medium was replaced with fresh serum-free medium, and cells were treated for 48 h with various components, including macrophage-conditioned media (CM), PKM2-IN-1 (MedChemExpress, USA), CCL1 (MedChemExpress, USA), and R243 (MedChemExpress, USA).

Techniques: Activation Assay, Over Expression, MANN-WHITNEY

Journal: PLoS ONE

Article Title: The Early Activation Marker CD69 Regulates the Expression of Chemokines and CD4 T Cell Accumulation in Intestine

doi: 10.1371/journal.pone.0065413

Figure Lengend Snippet: Expression of selected chemokine-related genes differentially expressed in CD69-activated B6 compared to CD69 −/− CD4 T cells analyzed by microarray.

Article Snippet: In one experiment mice were treated with chemokine antagonists (anti-CCL-1 Ab 148113 (cat. no. MAB845), anti-CXCL-10 Ab 134013 (cat. no. MAB466) and anti-CCL-19 Ab 87201 (cat no. MAB880) all purchased from R&D Systems, Wiesbaden, Germany).

Techniques: Expressing, Microarray

Journal: PLoS ONE

Article Title: The Early Activation Marker CD69 Regulates the Expression of Chemokines and CD4 T Cell Accumulation in Intestine

doi: 10.1371/journal.pone.0065413

Figure Lengend Snippet: CD4 T cells were enriched from the spleen of B6 or CD69 −/− mice and migration of these cells was analysed in vitro in transwell system. Medium alone or containing indicated concentrations of CCL-1, CXCL-10, CCL-5 and CCL-4 were loaded in the lower chamber of the transwell system, while CD4 T cells in the medium were loaded in the upper chamber. Cell number migrating from the upper chamber to the chemokine containing chamber trough polycarbonate membrane after 1 h at 37°C, 5% CO 2 was counted by flow cytometry. All the reactions were done in at least four repeats and average cell number per chemokine concentration per cell type was calculated. Results are presented as the number of cells migrating to the wells containing medium alone ( A ) or indicated concentration of CCL-1 ( B ), CXCL-10 ( C ), CCL-5 ( D ) or CCL-4 ( E ). Mean (± SEM) for at least four repeats per each chemokine concentration and cell type is presented. The lower graph set represents chemotactic indexes of B6 and CD69 −/− CD4 T cells for CCL-1 ( F) , CXCL-10 ( G) , CCL-5 ( H ) and CCL-4 ( I ). The chemotactic index was calculated as: the number of cells migrating to the media with chemokine divided by the average number of cells migrating to the media alone. *p≤0.05.

Article Snippet: In one experiment mice were treated with chemokine antagonists (anti-CCL-1 Ab 148113 (cat. no. MAB845), anti-CXCL-10 Ab 134013 (cat. no. MAB466) and anti-CCL-19 Ab 87201 (cat no. MAB880) all purchased from R&D Systems, Wiesbaden, Germany).

Techniques: Migration, In Vitro, Membrane, Flow Cytometry, Concentration Assay