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. Author manuscript; available in PMC: 2023 Mar 7.
Published in final edited form as: Cell Rep. 2023 Jan 23;42(1):112024. doi: 10.1016/j.celrep.2023.112024

Figure 5. ABS treatment causes conformation change of the p53 protein molecule.

Figure 5.

(A) Dot blot analysis of CP-A cell extracts using D01 antibody.

(B) The same as (A) but p53 (PAb 240) antibody was used. Staining was higher in cells treated with ABS than in control or 2-HOBA-treated cells (***p < 0.001; n = 3; Tukey’s multiple comparison).

(C) The same as (B) but EPC-2 cells were studied (ABS versus ABS+2-HOBA, ***p < 0.001; n = 3; Tukey’s multiple comparison).

(D) Native blue PAGE demonstrates misfolding and aggregation of p53 protein in CP-A cells collected after treatment with ABS. The graph shows the densitometric measurement of p53 aggregation detected by p53 (PAb 240) antibody (control versus ABS, ***p < 0.001; ABS versus 2-HOBA, ***p < 0.001; n = 3; Tukey’s multiple comparison). Aggregated p53 proteins are indicated by a red box. Protein expression in control cells was arbitrarily set at 1.

(E) Representative images show co-localization of p53 protein with ProteoStat-positive aggregates (scale bar, 5 μm). p53 was analyzed with p53(D0-1) and ProteoStat dye was used to identify protein aggregates in CP-A cells treated with ABS alone or in combination with 2-HOBA (n = 3).

(F) The same as (E) but PAb 240 antibody was used for detection of misfolded p53 protein (scale bar, 5 μm).

(G) The same as (F) but the p53 misfolding was analyzed in EPC-2 cells. All results are expressed as mean ± SD.