Fig 2: Immunohistochemistry of the four proteins in the colorectal tumors and matching normal tissue margins (control). Representative images of the expression of P504S, Bcl-2, p53 and Ki-67 in colorectal tumors and matching normal tissue margins are shown. P504S, a-methylacyl-CoA racemase; Bcl-2, B-cell lymphoma 2; p53, tumor protein p53.

Fig 3: Western blot analysis of the four proteins in the colorectal tumors and matching normal tissue margins (control). (A) Western blot analysis of the four proteins. (B) Expression levels of the four proteins normalized to ß-actin. *P<0.001. P504S, a-methylacyl-CoA racemase; Bcl-2, B-cell lymphoma 2; p53, tumor protein p53.

Fig 4: Effect of LMB-100 treatment on level of tumor cell proteins. (A) KLM1 cells were treated for 48 h with LMB-100 as per schema. Equal numbers of cells were lysed into a constant volume of lysis buffer and protein concentration assayed by standard colorimetric assay. (B) Panc02/chi-MSLN cells were treated with LMB-100 for 24 h as per schema. Protein concentration was assessed as described in (A). (C) KLM1 cells were treated as per schema and lysate assayed by RPPA for levels of the ubiquitinated forms of 49 protein analytes. “Reference” indicates control spots used to assess loading. (D) Immunoblot of lysates from (C) to assess level of Bcl-2. (E) KLM1 cells stably transfected with WT GFP and PEST-mCherry reporters driven off the same promoter through an IRES were treated with LMB-100 as per schema (Treat = start of LMB-100 treatment, ?M = change medium). Percentage of cells with fluorescence above background in red and green channels was assessed by flow cytometry. (F) KLM1 cells treated as shown in schema were lysed and protein levels assayed by RPPA for levels of 84 cancer-related proteins. (G) Immunoblot of lysates from treated cells.

Fig 5: LSDP5 reduces apoptosis in LO2 lipotoxicity Model cells. (A) Apoptosis was detected by Annexin V-FITC/PI apoptosis detection kit. (B) mRNA expression levels of Bax and Bcl-2 were measured by reverse transcription-quantitative polymerase chain reaction. (C and D) Protein expression levels of active-caspase-3, Bax and Bcl-2 were measured by western blot assay; ß-actin was used as a loading control and for normalization. *P<0.05, **P<0.01 and ***P<0.001 vs. Control; ^P<0.05 and ^^P<0.01 vs. Model. Bax, Bcl-2-associated X protein; Bcl-2, B-cell lymphoma-2; FITC, fluorescein isothiocyanate; LSDP5, lipid storage droplet protein 5; M, model; NC, negative control; PI, propidium iodide.