Figure 7.

Loss of NFKBIZ dysregulates the IL-6 axis

(A and B) GSVA scores for TGFβ or WNT signaling as indicated for different ROIs.

(C) Expression in ROIs for SOD2.

(D) GSVA scores for different ROIs for hallmark interferon alpha response.

(E and F) Expression in ROIs for NFKBIZ (E) and GADD45B (F).

(G) GSVA scores for different ROIs for GOresponse to interleukin 6. (H) Expression in ROIs for IL6.

(I–K). Expression of NFKBIZ (I), IL6 (J), and GADD45B (K) in GSE32537.

(L–N) Expression of NFKBIZ (L), IL6 (M), and GADD45B (N) in GSE169500. Statistical comparisons are relative to control alveolar septae. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001 by Wilcoxon test with Benjamini-Hochberg multiple test correction.

(O) Representative multiplexed RNA in situ hybridization for SFTPC and NFKBIZ expression in control alveolar septae and IPF distal alveolar septae. Scale bar, 100 μm; inset scale bar, 10 μm.

(P–R) Type 2 alveolar epithelial cells were treated with TGFβ or vehicle control as indicated for 24 hr. Relative gene expression of NFKBIZ (P), IL6 (Q), and GADD45B (R) determined by qRT-PCR and analyzed using ΔΔCt method. Data are mean ± SD; n = 7 across three independent experiments. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001 by two-way ANOVA with Tukey’s multiple comparison test.

(S–U) Type 2 alveolar epithelial cells were transfected with NFKBIZ targeting siRNA or control siRNA for 48 hr followed by treatment for 24 hr with TGFβ or vehicle control as indicated. (S) Relative gene expression of IL6 measured as in (Q). (T) IL-6 protein in conditioned media quantified by ELISA. (U) Relative gene expression of GADD45B measured as in (T). Data are mean ± SD; n = 6 across three independent experiments. ^∗p < 0.05, ^∗∗p < 0.01, ^∗∗∗p < 0.001, ^∗∗∗∗p < 0.0001 by two-way ANOVA with Tukey’s multiple comparison test.