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. 2019 Aug 10;13(10):2142–2159. doi: 10.1002/1878-0261.12552

Figure 4.

Figure 4

NF‐kB/p65‐mediated regulation of progranulin expression in human normal and cancerous colonic cell lines. (A) Representative western blotting showing progranulin expression in HCEC‐1CT cells stimulated or not with IL‐6, IL‐22, TNF‐α, and IL‐17A (all used at 25 ng·mL−1) for 8 h. β‐actin was used as a loading control. One of four representative experiments in which similar results were obtained is shown. Right inset. Quantitative analysis of progranulin/β‐actin protein ratio in total extracts of HCEC‐1CT cells stimulated or not with IL‐6, IL‐22, TNF‐α, and IL‐17A for 8 h, as measured by densitometry scanning of western blots. Values are expressed in arbitrary units and are the mean ± SEM of four experiments. Differences among groups were compared using one‐way analysis of variance (ANOVA) followed by Tukey's post hoc test. TNF‐α‐stimulated cells vs untreated cells, ***P < 0.001. (B) Representative western blotting showing p‐STAT3 Tyr705, p‐NF‐kB/p65 Ser536, p‐p38, and p‐ERK expression in HCEC‐1CT cells stimulated or not with IL‐6, IL‐22, TNF‐α, and IL‐17A (all used at 25 ng·mL−1) for 15 min (left panels), 30 min (middle panels), and 8 h (right panels). β‐actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown. (C) Representative western blotting showing progranulin, p‐NF‐kB/p65 Ser536, p‐p38, and p‐ERK expression in HCEC‐1CT cells either left untreated or stimulated with TNF‐α (used at 25 ng·mL−1) for 8 h in the presence of NF‐kB/p65 inhibitor (BAY 11‐7082), p38 inhibitor (SB202190), ERK inhibitor (PD98059), or DMSO (vehicle) as indicated. β‐actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown. (D). Representative western blotting showing progranulin, p‐NF‐kB/p65 Ser536, p‐p38, and p‐ERK expression in HT‐29 cells cultured for 8 h in the presence of NF‐kB/p65 inhibitor (BAY 11‐7082), p38 inhibitor (SB202190), ERK inhibitor (PD98059), or DMSO (vehicle) as indicated. β‐actin was used as a loading control. One of three representative experiments in which similar results were obtained is shown.