Fig 1: Exploring the SASP cytokines uniquely high-expressed in SenV.(A) The expression heatmap of cytokine genes, which showed the highest expression in SenV compared to NC and SenC. Cell senescence was induced by treating cells once with Vem (10 μM) or CDDP (2 μM) and RNA-seq was performed on day 7 after treatment. CCL2, TIMP2, and NGFR were chosen for further verification. (B) The relative mRNA levels of NGFR, CCL2, and TIMP2 in senescent A375 cells induced by a single treatment with Vem (10 μM), continuous treatment with Vem (0.5 μM), or single treatment with CDDP (2 μM). (C–D) Relative mRNA levels of NGFR, CCL2, and TIMP2 in senescent M14 and A875 cells induced by continuous Vem treatment. (E–F) The relative secretion levels of CCL2 and TIMP2 in Sen CM induced by continuous Vem treatment, evaluated by ELISA. *P < 0.05, **P < 0.01 vs. Vehicle (n = 3). NS means not significant.
Fig 2: The cytokines CCL2, TIMP2, and NGFR display resistance to Vem.CCL2, TIMP2, and NGFR were diluted and added into 10% FBS culture medium with or without Vem. The effects of these cytokines on cell proliferation and Vem sensitivity were evaluated by clone formation. When obvious clones could be observed, the cells were fixed and stained. Due to different growth rates, the NC and Vem groups might be stained at different times. Horizontal comparison revealed that these cytokines alone had negligible or weak effects on cell proliferation, but they generally displayed resistance to Vem. Similar results were observed in 3 independent experiments. The red quadrangles contain the control and experiment groups to facilitate comparison.>
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