Extended Data Fig. 2: (R)-NbALFA does not affect protein degradation.
Parental cells (U2OS with (R)NbALFA-mEmerald stably integrated into the genome), U2OS-ALFA-CKAP4, and U2OS-ALFA-LMNA cell lines were treated with 50 µg/ml CHX for 0, 12 and 24 h. a,c, Total protein from these cells was extracted using RIPA lysis buffer, separated by SDS-PAGE, and immunoblotted with anti-CKAP4, anti-LMNA and anti-GAPDH antibodies. P53 as positive controls. b,d, Quantitative analysis of CKAP4 (b) and LMNA (d) levels, normalized to GAPDH. The average values from three biologically independent experiments were calculated using Fiji software, with CKAP4 and LMNA amounts normalized to the group without CHX treatment. Data are expressed as the mean ± s.e.m. Significance was calculated by two-tailed Student’s t-test CHX: cycloheximide.
