Journal: Nature

Article Title: Probing condensate microenvironments with a micropeptide killswitch

doi: 10.1038/s41586-025-09141-5

Figure Lengend Snippet: a . Live cell fluorescence microscopy images of U2OS cells expressing ectopic mEGFP-fsHMGB1 variants and RFP-FIB1. The cell nucleus is highlighted with a dashed white line contour. Scale bar: 5 µm. b . (left) Model of fsHMGB1 and sequences of the KS variants within the tested mEGFP-HMGB1 proteins. (right) Quantification of circularity of nucleoli in cells expressing ectopic mEGFP-HMGB1 KS variants. P -values from Dunnett’s multiple comparison test versus fsHMGB1-full length after one-way ANOVA. P ( Δkillswitch ) = <0.0001, P ( F-to-E&D ) = < 0.0001, P ( F-to-A ) = 0.0009, P ( F-to-G ) = < 0.0001, P ( 0F ) = 0.0972, P ( F12A ) = 0.0003, P ( F13A ) = > 0.9999, P ( ΔKS-3F ) = < 0.0001, P ( 11G3F3G ) = < 0.0001, P ( C16-to-A ) = 0.0346, P ( M-to-E&D ) = < 0.0001. ****: P < 10 −4 , ***: P < 10 −3 , * : P < 0.05. c . FRAP of the indicated mEGFP-HMGB1 variants. The quantification of the corresponding mobile/immobile fraction ratios are shown in Fig. . Data are mean ± s.d. d . Cell viability of U2OS cell expressing mEGFP-HMGB1 proteins. Data are mean ± s.d. P -values from one-way ANOVA Dunnett’s multiple comparison test versus fsHMGB1-full length. (Left): P ( Full length ) = <0.0001, P ( Δkillswitch ) = 0.5987, P ( F-to-D&E ) = 0.0320, P ( M-to-D&E ) = < 0.0001, P ( C-to-A ) = < 0.0001; (Middle): P ( Full length ) = <0.0001, P ( Δkillswitch ) = 0.9975, P ( F-to-D&E ) = 0.8919, P ( F-to-A ) = 0.9967, P ( F-to-G ) = < 0.5244. (Right): P ( Full length ) = 0.0046, P ( Δkillswitch ) = 0.2873, P ( F-to-G ) = 0.4834, P ( ΔKS-3F ) = 0.4818, P ( 0F ) = 0.6900, P ( 11G3F3G ) = 0.4578. n = three (except two for F-to-D&E from the middle plot and ΔKS-3F from the right plot) biologically independent experiments. e . Representative live cell fluorescence microscopy images of A673 cells expressing EGFP-NPM1-WT and -KS variants 24 h after doxycycline induction. The cell nucleus is highlighted with a dashed white line contour. The experiments were repeated independently twice with similar results. Scale bar: 5 µm. f . Quantification of the mobile and immobile fractions of EGFP-NPM1 proteins. Data are mean ± s.d. n = 15 cells for all samples from two biologically independent experiments. P -values from Dunnett’s multiple comparisons test versus EGFP-NPM1-WT after one-way ANOVA. P ( KS ) = < 0.0001, P ( KS_F-to-G ) = 0.76, P ( KS_F-to-A ) = 0.03, P ( KS_0F ) = 1.00. g . Mean GFP fluorescence of the bleached area. Data are mean ± s.d. n = 15 cells for all samples from two biologically independent experiments. P -values from Dunnett’s multiple comparisons test versus EGFP-NPM1-WT after one-way ANOVA.

Article Snippet: U2-OS (ATCC, HTB-96) HEK293T (ATCC, CRL-3216), HCT-116 (ATCC, CCL-247), MCF7 (ATCC, HTB-22), C2C12 (ATCC, CRL-1772), Lenti-X 293T (Takara Bio, 632180) and A673 (gifted by H. Kovar; CLS, 300454) cell lines were cultured in DMEM GlutaMAX (Gibco, 31966047).

Techniques: Fluorescence, Microscopy, Expressing, Comparison

Journal: Nature cancer

Article Title: Distinct CDK6 complexes determine tumor cell response to CDK4/6 inhibitors and degraders

doi: 10.1038/s43018-021-00174-z

Figure Lengend Snippet: a, A673 and TC-71 cells were transfected with non-targeting control or siCDK4 or siCDK6 for 72 hr. Lysates were immunoblotted with the indicated antibodies. b, Relationship between CDK4 and CDK6 expression (CCLE RNA-seq) and DepMap CRISPR–Cas9 single-gene knockout scores (CERES; 20Q1 public dataset). All expression values are in log 2 (TPM +1). Cell lines harboring COSMIC hotspot mutations to RB1 are annotated in orange. P -values were calculated based on linear regression analysis.

Article Snippet: A673 and TC-71 were provided by Dr. Christine A. Pratilas (Johns Hopkins).

Techniques: Transfection, Control, Expressing, RNA Sequencing, CRISPR, Gene Knockout