Fig. 1: RNAi screens identify senolytic targets.
From: COPI vesicle formation and N-myristoylation are targetable vulnerabilities of senescent cells
a, Experimental design for the RNAi screens to identify senolytic targets. b, Right: quantification of cell survival of senescent (4OHT) and control (DMSO) IMR90 ER:RAS cells three days post-transfection with BCL2L1 siRNA (n = 3). Left: representative DAPI-stained immunofluorescence (IF) images. Scale bar, 100 µm. c, Results of the primary siRNA screen for senolytic targets in OIS. Normalized cell counts are shown as mean B-score, reflecting counts normalized to account for plate positional effects using the B-scoring method. A candidate was considered a hit if the B-score in ≥2 replicates was <−3. d, Summary of the siRNA screen for senolytic targets in OIS, genes in the same pathway are indicated in bold. e, Re-test of OIS screen candidates. A candidate was considered a hit if the change in % cell survival was >20 with siRNAs in ≥2 replicates. f, Percentage cell survival in the context of OIS (4OHT) and control (DMSO) cells (n = 3). The data represent the deconvolution of values shown in e. g, Right: quantification of cell survival of doxorubicin-induced senescent (Doxo) and control (DMSO) IMR90 cells three days post-transfection with BCL2L1 siRNA (n = 3). Left: representative DAPI IF images. Scale bar, 100 µm. h, Results of the primary siRNA screen for senolytic targets in doxorubicin-induced senescence. Normalized cell counts are shown as mean B-score. A candidate is considered a hit if the B-score was <−3 in ≥2 replicates. i, Summary of the siRNA screen for senolytic targets in doxorubicin-induced senescence. j, Re-test of TIS screen candidates. A candidate was considered a hit if the change in % cell survival was >15 with siRNAs in ≥2 replicates. k, Percentage cell survival of doxorubicin-induced senescence (Doxo) and control (DMSO) cells (n = 6 for DMSO- and 4OHT-treated cells, n = 3 for BCLXL siRNA transfected cells). Data represent the deconvolution of values shown in j with additional replicates. l, Common pathways identified in the siRNA screen for senolytic targets. Data in b, f, g and k are presented as mean ± s.d. (unpaired, two-tailed Student’s t-test). n represents independent experiments in b, f, g, k. Data in c and h is representative of three replicates. Data are presented as percentage cell survival in control cells versus the difference in cell survival between control and senescent cells in e and j. Source numerical data are available as source data.
