Figure 2: Polyinosinic:polycytidylic acid (poly[I:C]) induces pyroptosis and NLRP3 inflammasome activation in outer root sheath (ORS) cells. | Scientific Reports

Figure 2: Polyinosinic:polycytidylic acid (poly[I:C]) induces pyroptosis and NLRP3 inflammasome activation in outer root sheath (ORS) cells.

From: Double-stranded RNA induces inflammation via the NF-κB pathway and inflammasome activation in the outer root sheath cells of hair follicles

Figure 2

ORS cells were treated with poly(I:C) at the indicated concentrations for 24 h. (a) Cell viability was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. (b) Pyroptosis was determined by lactate dehydrogenase activity. (c) IL-1β secretion in supernatants (SN) was assessed using enzyme-linked immunosorbent assay (ELISA). (d) Relative mRNA expression of IL-1β, NLRP3, ASC, and caspase-1 was determined by quantitative reverse transcription polymerase chain reaction. (e) Expression levels of NLRP3, pro-caspase-1, and IL-1β in the cell lysates (CL) were assessed by Western blotting. Actin was used as a loading control. Expression levels of active caspase-1 p20 subunit and mature IL-1β p17 in SN were determined by Western blotting. (f) Co-localization of NLRP3 (red) and ASC (green) was detected by immunofluorescence staining. Nuclei were counterstained with 4,6-diamidino-2-phenylindole (blue). Bars = 20 μm. Data are presented as mean ± standard error of the mean (SEM). Data were analyzed by one-way ANOVA (a–c,e) or Student’s t-test (d) (n = 3, *p < 0.05, **p < 0.01). Cropped blots were used in this figure and full-length blots are presented in Supplementary Figure 4.

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