Fig. 6. Senescent cells have a reduced capacity to form aggresomes.
a, Cycling and senescent HFF-1 cells were treated with MG132 and stained as indicated. The number of passages (p) and total number of days in culture (d) are indicated. b, Quantitation of aggresome formation in Ki67 positive/negative cycling and senescent HFF-1 cells. c, Extracts from cycling and senescent HFF-1 cells treated with or without MG132 were probed as indicated. α-tubulin was used as the loading control. d, Cycling HFF-1 cells were treated with control (GL2), CP110 or CEP290 siRNAs, treated with MG132 and stained as indicated. e, Quantitation of aggresome formation in cycling HFF-1 cells treated as in d. f, Senescent HFF-1 cells were transfected with GFP–CP110 and treated with MG132. Cells were stained as indicated. g, Senescent HFF-1 cells were transfected with GFP–CP110, GFP–CP110-R586A,L588A or GFP–CP110-Δ67-82 and treated with MG132. Cells were stained for pHSP27. h, Quantification of senescent HFF-1 cells treated as in g that formed an aggresome. For b, e and h, data are displayed as the mean ± s.d., n = 3 independent experiments, and P values were calculated by two-tailed unpaired Student’s t-test; ****P 0.0001, ***P 0.001. Scale bars, 2 μm (insets of a,d,f,g) or 10 μm (a,d,f,g). Unprocessed immunoblots, numerical data and P values are provided as source data.