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. 2022 Mar 7;28(10):2167–2179. doi: 10.1158/1078-0432.CCR-21-3185

Figure 3.

Figure 3. Senescence. A, Study of the senescence-like phenotype in HER2+ BT474 cell line, and in HER2low MCF7 cell line. The effects of treatments with palbociclib and PFHPert were studied at EOT (72 hours) and after additional 3 days of WO with fresh medium. Morphologic changes after WO were shown in BF images on the left-sided panels. In the right-sided panels the SA β-gal enzyme activation in the same conditions was shown. For the complete panel of treatments in BT474, MCF7, and MDA-MB-361 see Supplementary Fig. S6. B, Representative images of morphologic changes in MCF7 cell treated with palbociclib for 72 hours. C, Evaluation of SA β-gal activation at baseline (untreated cells), during (6 hours), at the end (EOT), and after treatment (3 days WO) in BT474 (graph on the left), MDA-MB-361 (graph on the middle), and MCF7 (graph on the right). The comparison between untreated cells (white bars), palbociclib alone (yellow bars), and PFHPert (red bars) was reported. Staining was performed with a colorimetric kit and detection of % of SA β-gal–positive cells was evaluated by Amnis Image Stream Flow Cytometry Technology (Merck). Modulation of p53-p21-MDM2 axis and Cyclin D1 during treatment with palbociclib and PFHPert in BT474 (D) and MCF7 (E). Data are from two or three replicates but only one representative blot is showed. Values were calculated after correction for the loading marker and normalization by the control sample in DMSO. Bars represent the mean ± SEM. NT, untreated cells; h, hours.

Senescence. A, Study of the senescence-like phenotype in HER2+ BT474 cell line, and in HER2low MCF7 cell line. The effects of treatments with palbociclib and PFHPert were studied at EOT (72 hours) and after additional 3 days of WO with fresh medium. Morphologic changes after WO were shown in BF images on the left-sided panels. In the right-sided panels the SA β-gal enzyme activation in the same conditions was shown. For the complete panel of treatments in BT474, MCF7, and MDA-MB-361 see Supplementary Fig. S6. B, Representative images of morphologic changes in MCF7 cell treated with palbociclib for 72 hours. C, Evaluation of SA β-gal activation at baseline (untreated cells), during (6 hours), at the end (EOT), and after treatment (3 days WO) in BT474 (graph on the left), MDA-MB-361 (graph on the middle), and MCF7 (graph on the right). The comparison between untreated cells (white bars), palbociclib alone (yellow bars), and PFHPert (red bars) was reported. Staining was performed with a colorimetric kit and detection of % of SA β-gal–positive cells was evaluated by Amnis ImageStreamX Flow Cytometry Technology (Merck). Modulation of p53-p21-MDM2 axis and Cyclin D1 during treatment with palbociclib and PFHPert in BT474 (D) and MCF7 (E). Data are from two or three replicates but only one representative blot is showed. Values were calculated after correction for the loading marker and normalization by the control sample in DMSO. Bars represent the mean ± SEM. NT, untreated cells; h, hours.