Fig. 2: COPI is a vulnerability of senescent cells.
From: COPI vesicle formation and N-myristoylation are targetable vulnerabilities of senescent cells
a, Right: percentage cell survival of the indicated senescent (4OHT) and control (DMSO) IMR90 ER:RAS cells (n = 4, IMR90 Vector+4OHT; n = 5, other groups). Left: representative DAPI-stained IF images. Scale bar, 100 µm. b, Crystal violet staining. An image representative of three independent experiments is shown. c, Senolytic activity of COPB2 in doxorubicin-induced senescence in IMR90 cells (n = 4 shCOPB2.1, n = 5 other shRNAs). d, Senolytic activity of COPG1 depletion during OIS in IMR90 ER:RAS cells (n = 5, IMR90 Vector+4OHT; n = 6, other groups). e, Schematic outlining the strategy of co-culture senolytic testing of COPB2 siRNAs. f, Right: percentage survival in a co-culture experiment of IMR90 green fluorescent protein (GFP) ER:RAS with IMR90 Cherry cells transfected with the indicated siRNAs. Cell numbers were determined from counts of mCherry or GFP-positive cells detected by IF (n = 3). Left: representative IF images. Scale bar, 100 µm. g, Representative images of IMR90 ER:RAS cells seven days post addition of 4OHT and stained for Senescence-Associated (SA)-β-Gal activity 72 h after treatment with 2.5 µM GCA (n = 3). Quantification is shown in Extended Data Fig. 2a. Scale bar, 100 µm. h, Dose–response curves for the senolytic effect of the GBF1 inhibitor GCA in the context of OIS (n = 6). Red, IMR90 ER:RAS +4OHT; Black, IMR90 vector +4OHT; Blue, IMR90 ER:RAS +DMSO. i, Percentage survival of control cells (RAS DMSO) and oncogene-induced senescent cells (RAS 4OHT) transduced with vectors and treated with ABT-263, GCA or BFA (n = 5 for GCA/BFA; n = 3, ABT-263). j, Caspase-3/7 activity in control (DMSO) or senescent (4OHT) cells after treatment with DMSO or 2.5 µM GCA seven days after senescence induction (n = 3). k, Senolytic activity of GCA in senescence induced by irradiation (n = 3), bleomycin and DMSO (n = 6), and doxorubicin (n = 4). Data are presented as mean ± s.d. Comparisons to the corresponding DMSO-treated cells (grey bars) with two-way analysis of variance (ANOVA). l, Right: percentage cell survival of p16INK4a positive and negative cells in PBECs after treatment with GCA or vehicle (DMSO). Left: representative p16INK4a (green)-stained IF images. Scale bar, 50 μm. n = 3. Data in all figures are presented as mean ± s.d. n represents independent experiments throughout the figure. Unpaired, two-tailed, Student’s t-test was used unless otherwise stated. Source numerical data are available as source data.
