Fig. 3
From: N-acetylcysteine stimulates organelle malfunction in endometriotic cells via IFN-gamma signaling
Effects of NAC, IFN-ɣ and IFN-ɣ receptor antagonist (Anta) on proliferation and IRE1 signaling in 12Z cells (a) xCELLigence analysis of 12Z cells individually and in combination with NAC, IFN-ɣ, Anta. Blocking IFN-ɣ signaling reduced IFN-ɣ and IFN-ɣ+NAC mediated antiproliferative effects (b–j) Ki-67 (green) and p-IRE1 (red) immunofluorescence labeling of 12Z cells. NAC, IFN-ɣ, and IFN-ɣ+NAC treatments decreased Ki-67 labeling compared to Anta including groups, significantly while p-IRE1 expression was significantly higher in IFN-ɣ-treated cells compared to the IFN-ɣ+Anta and IFN-ɣ+NAC + Anta groups. NAC treatment also increased p-IRE1 levels compared to the IFN-ɣ+Anta and IFN-ɣ+NAC + Anta groups. DAPI (blue) Scale bars: 75 μm (630x). (n = 3) Statistical differences between groups are indicated by bars and are significant at *p < 0.05, and **p ≤ 0.01 levels. (k) Representative western blotting analysis of p-IRE1 levels were relatively highest in IFN-ɣ treated cells, followed by NAC, IFN-ɣ+NAC, IFN-ɣ+NAC + Anta, IFN-ɣ+Anta, and the control groups. The bar graph shows that IRE1 protein level normalized to the β-actin protein (n = 3). Agilent RTCA Software (v 2.0) was used to create xCELLigence images, and GraphPad Prism (v 10.4.1) was used to create statistical graphics. ImageJ (v 1.54 g) was used to create immunofluorescence images and western blotting images.
