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. 2022 Feb 21;18(5):1912–1932. doi: 10.7150/ijbs.68484

Figure 8.

Figure 8

High doses of palmitate aggravated the lipotoxicity of CAF-CM- treated CRC cells. A-B, Quantification of diacylglycerol (DG; A) and triacylglycerol (TG; B) by lipidomics. C, Growth curves of DLD1 cells treated with the indicated compounds. IncuCyte ZOOM software was used to process the image data for phase object confluence. n=3. D, C16:0 induced apoptotic cell death in DLD1 cells. Cell viability assay of DLD1 cells treated with different concentrations of C16:0 (0, 50 and 100 µM) for 48 h and then detected by the annexin-V/PI assay. Apoptotic cells were identified by propidium iodide (PI) and annexin V staining. n=3. E, Reactive oxygen species (ROS) levels in DLD1 cells with or without CAF-CM incubation and SCD inhibitor treatment were detected by flow cytometry. n = 3. F, SCD inhibitor treatment increased cellular sensitivity to C16:0 in DLD1 cells. Cell viability was assessed by annexin-V/PI assay. n=3. G, BIP (GRP78 BiP) protein levels in DLD1/HCT116 cells incubated with CAF-CM or after C16:0 treatment (50 µM, ~6 h) was detected by Western blotting. Anti-BIP antibody (1:1000 dilution; Abcam, Cambridge, MA, USA), n=3. H, Relative protein level of BIP in DLD1 cells. I, DLD1 cells incubated with CAF-CM and then treated with C16:0 were detected by oil red/hematoxylin staining. The cells were imaged using an inverted microscope. n = 3. J, Cell viability assay of the indicated DLD1 cells treated with different concentrations of C16:0 (0, 50, 100 and 200 µM) for 48 h and then detected by the annexin-V/PI assay. Apoptotic cells were identified by propidium iodide (PI) and annexin V staining. n=3. Bars, mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001.