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. 2025 Jul 26;34:102138. doi: 10.1016/j.mtbio.2025.102138

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© 2025 The Authors

This is an open access article under the CC BY-NC license (http://creativecommons.org/licenses/by-nc/4.0/).

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Fig. 5 — Evaluation of the molecular mechanisms underlying the effects of different microsphere formulations on chondrocyte senescence under mechanical stress. A) Relative expression levels of ETS2 mRNA among the control, HMs, and Lipo@HMs groups. B) Quantification of RSK gene expression across experimental groups. C) Quantification of TNIK gene expression across experimental groups. D) Analysis of c-Fos mRNA levels among the control, HMs, and Lipo@HMs groups. E) Quantification of C/EBPβ gene expression across experimental groups. F) Western blot showing protein expression of C/EBPβ across four experimental groups, including control, HMs, HMs + P38 inhibitor, and Lipo@HMs. G) Western blot analysis of c-Fos levels among the control, HMs, HMs + ERK inhibitor, and Lipo@HMs groups. H) Quantitative analysis of c-Fos by Western blotting. I) Quantitative analysis of C/EBPβ by Western blotting. J) Western blot results of ERK and P38 phosphorylation under mechanical stress. K) The phosphorylation level of ERK was assessed by the ratio of P-ERK to total ERK. L) The phosphorylation level of P38 was assessed by the ratio of P-P38 to total P38. M) Expression levels of cellular senescence markers P21 and P16INK4a in different treatment groups. N) Quantitative analysis of P21 by Western blotting. O) Quantitative analysis of P16INK4a by Western blotting. P) Flow cytometric analysis of apoptosis across different treatment groups. Q) Quantitative analysis of apoptosis in chondrocytes treated with HMs, Lipo@HMs, and Res@Lipo@HMs under mechanical stress. R) Schematic representation of the signaling pathways involved in mechanical stress–induced chondrocyte senescence (n = 3 for each group); (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ns: not significant).

Fig. 5 — Evaluation of the molecular mechanisms underlying the effects of different microsphere formulations on chondrocyte senescence under mechanical stress. A) Relative expression levels of ETS2 mRNA among the control, HMs, and Lipo@HMs groups. B) Quantification of RSK gene expression across experimental groups. C) Quantification of TNIK gene expression across experimental groups. D) Analysis of c-Fos mRNA levels among the control, HMs, and Lipo@HMs groups. E) Quantification of C/EBPβ gene expression across experimental groups. F) Western blot showing protein expression of C/EBPβ across four experimental groups, including control, HMs, HMs + P38 inhibitor, and Lipo@HMs. G) Western blot analysis of c-Fos levels among the control, HMs, HMs + ERK inhibitor, and Lipo@HMs groups. H) Quantitative analysis of c-Fos by Western blotting. I) Quantitative analysis of C/EBPβ by Western blotting. J) Western blot results of ERK and P38 phosphorylation under mechanical stress. K) The phosphorylation level of ERK was assessed by the ratio of P-ERK to total ERK. L) The phosphorylation level of P38 was assessed by the ratio of P-P38 to total P38. M) Expression levels of cellular senescence markers P21 and P16INK4a in different treatment groups. N) Quantitative analysis of P21 by Western blotting. O) Quantitative analysis of P16INK4a by Western blotting. P) Flow cytometric analysis of apoptosis across different treatment groups. Q) Quantitative analysis of apoptosis in chondrocytes treated with HMs, Lipo@HMs, and Res@Lipo@HMs under mechanical stress. R) Schematic representation of the signaling pathways involved in mechanical stress–induced chondrocyte senescence (n = 3 for each group); (∗P < 0.05, ∗∗P < 0.01, ∗∗∗P < 0.001, ns: not significant).