Journal: bioRxiv

Article Title: IL-1 instructs para-bronchial cuff fibroblasts to organize lung antibody secreting cell niches during continued antigen exposure

doi: 10.64898/2026.01.09.698642

Figure Lengend Snippet: A. CellChat analysis on the chronic HDM lung dataset shows strong IL-1 interactions between macrophages/DCs to fibroblasts. B. UMAP of integrated lung macrophages and DCs following 5 weeks of HDM challenges C. DotPlot showing expression of Il1a and Il1b expression on lung macrophage and DC subsets D. Schematic of in vitro stimulation of isolated lung fibroblasts with recombinant IL-1 E. BAFF, CXCL12 and IL-6 secretion by IL-1 stimulated lung fibroblasts F. UMAP of integrated lung fibroblasts following 5 weeks of HDM challenges G. DotPlot showing expression of ASC survival factors Tnfsf13b , Cxcl12 and Il6 , and Il1r1 and Pdgfra expression on lung fibroblast subsets H. Experimental set-up for temporal deletion of IL-1 signaling in Pdgfra+ fibroblasts during the chronic HDM protocol I. To L. Quantification of lung ASCs in Il1r1 fl/+ x Pdgfra CreERT2Tg/+ and Il1r1 fl/− x Pdgfra CreERT2Tg/+ mice. I. showing total lung ASCs J. Quantification of IgG1 + lung ASCs K. Quantification of IgA + lung ASCs L. Quantification of IgM + lung ASCs M. Confocal analysis of ASC positioning into the para-bronchial cuff in Il1r1 fl/+ x Pdgfra CreERT2Tg/+ and Il1r1 fl/− x Pdgfra CreERT2Tg/+ mice (scale bars: 700μm for overview images and 300μm for ROIs). ROIs show example of ASC track from TLO to calculate ASC-TLO distance and CD138 intensity profile. The dotted line on WT ROI (Il1r1 fl/+ x Pdgfra CreERT2Tg/+ ) was used for distance measurement. N. Quantification of ASC distance from TLO in Il1r1 fl/+ x Pdgfra CreERT2Tg/+ and Il1r1 fl/− x Pdgfra CreERT2Tg/+ mice O. CD138 intensity profile along ASC tracks from TLO in Il1r1 fl/+ x Pdgfra CreERT2Tg/+ and Il1r1 fl/− x Pdgfra CreERT2Tg/+ mice, showing that ASCs tend to reside more within the TLO in the absence of IL-1R signaling on Pdgfra+ fibroblasts. Blue boxes indicate TLO areas along the ASC track. ROIs for the tracks are indicated in M P. Total IgG1, IgA and IgM BAL levels in Il1r1 fl/+ x Pdgfra CreERT2Tg/+ and Il1r1 fl/− x Pdgfra CreERT2Tg/+ mice For E, I-L, N, P, bars represent median with each symbol representing 1 replicate. P values are for Mann-Whitney U test (*P<0,05, **P<0,01, ***P<0,001). Data are representative of 3 independent experiments.

Article Snippet: IL1r1 −/− mice (strain #003245) were crossed to PdgfraCreERT2 Tg/+ mice, and Il1r1 −/+ x Pdgfra CreERT2Tg/+ (strain #032770) mice were crossed to Il1r1 fl/fl (strain #028398, purchased from Jackson Laboratory) to obtain Il1r1 fl/− x Pdgfra CreERT2Tg/+ mice.

Techniques: Expressing, In Vitro, Isolation, Recombinant, MANN-WHITNEY

Journal: Scientific reports

Article Title: GPER signalling in both cancer-associated fibroblasts and breast cancer cells mediates a feedforward IL1β/IL1R1 response.

doi: 10.1038/srep24354

Figure Lengend Snippet: Figure 2. E2 and G-1 induce IL1R1 expression in SkBr3 and MCF-7 breast cancer cells. 10 nM E2 (A) and 100 nM G-1 (B) induce the mRNA expression of IL1R1, as evaluated by real-time PCR. Data obtained in three independent experiments performed in triplicate were normalized to 18S expression and shown as fold changes of IL1R1 expression upon E2 and G-1 treatments respect to cells exposed to vehicle (−). (◼) p < 0.05 for cells receiving treatments versus vehicle. Evaluation of IL1R1 protein expression in SkBr3 (C,D) and MCF-7 cells (E,F) treated with 10 nM E2 and 100 nM G-1, as indicated. β -actin serves as a loading control. Results shown are representative of at least two independent experiments.

Article Snippet: Equal amounts of whole protein extract were resolved on a 10% SDS-polyacrylamide gel, transferred to a nitrocellulose membrane (Amersham Biosciences, GE Healthcare, Milan, Italy), probed overnight at 4 °C with antibodies against IL1β (R&D Systems, Inc. Celbio, Milan, Italy), IL1R1 (OriGene Technologies, TEMA ricerca srl, Bologna, Italy), GPER (N-15), PGE synthase (S-16) and β -actin (C-2) all purchased from Santa Cruz Biotechnology, DBA, Milan, Italy).

Techniques: Expressing, Real-time Polymerase Chain Reaction, Control

Journal: Scientific reports

Article Title: GPER signalling in both cancer-associated fibroblasts and breast cancer cells mediates a feedforward IL1β/IL1R1 response.

doi: 10.1038/srep24354

Figure Lengend Snippet: Figure 3. GPER mediates the up-regulation of IL1R1 expression by E2 and G-1 in SkBr3 and MCF-7 breast cancer cells. (A) The up-regulation of IL1R1 protein levels upon treatment for 8 h with 10 nM E2 and 100 nM G-1 is abrogated transfecting SkBr3 cells for 24 h with shGPER. (B) Efficacy of GPER silencing. (C) The induction of IL1R1 protein expression observed treating SkBr3 cells for 8 h with 10 nM E2 and 100 nM G-1 is abolished in the presence of 100 nM GPER antagonist G-15. (D) The up-regulation of IL1R1 protein levels upon treatment for 8 h with 10 nM E2 and 100 nM G-1 is abrogated transfecting MCF-7 cells for 24 h with shGPER. (E) Efficacy of GPER silencing. (F) The induction of IL1R1 protein expression observed treating MCF-7 cells for 8 h with 10 nM E2 and 100 nM G-1 is abolished in the presence of 100 nM GPER antagonist G-15. IL1R1 protein levels in SkBr3 cells treated for 8 h with 10 nM E2 (G) and 100 nM G-1 (H) alone or in combination with 1 μM EGFR inhibitor AG1478 (AG), 1 μM MEK inhibitor PD98059 (PD), 1 μM PKC inhibitor GF109203X (GF), 1 μM PI3K inhibitor LY294,002 (LY), 1 μM PKA inhibitor H89 and 1 μM p38 MAPK inhibitor SB 203580 (SB). IL1R1 protein levels in MCF-7 cells treated for 8 h with 10 nM E2 (I) and 100 nM G-1 (J) alone or in combination with 1 μM EGFR inhibitor AG, 1 μM MEK inhibitor PD, 1 μM PKC inhibitor GF, 1 μM PI3K inhibitor LY, 1 μM PKA inhibitor H89 and 1 μM p38 MAPK inhibitor SB. β -actin serves as a loading control. Results shown are representative of at least two independent experiments.

Article Snippet: Equal amounts of whole protein extract were resolved on a 10% SDS-polyacrylamide gel, transferred to a nitrocellulose membrane (Amersham Biosciences, GE Healthcare, Milan, Italy), probed overnight at 4 °C with antibodies against IL1β (R&D Systems, Inc. Celbio, Milan, Italy), IL1R1 (OriGene Technologies, TEMA ricerca srl, Bologna, Italy), GPER (N-15), PGE synthase (S-16) and β -actin (C-2) all purchased from Santa Cruz Biotechnology, DBA, Milan, Italy).

Techniques: Expressing, Control

Journal: Scientific reports

Article Title: GPER signalling in both cancer-associated fibroblasts and breast cancer cells mediates a feedforward IL1β/IL1R1 response.

doi: 10.1038/srep24354

Figure Lengend Snippet: Figure 5. PTGES protein expression in SkBr3 (A–C) and MCF-7 (D–F) cells treated with 10 ng/ml IL1β alone or treated for 8 h with 10 nM E2 or 100 nM G-1 and then exposed to 10 ng/ml IL1β , as indicated. Protein levels of PTGES in SkBr3 (G,H) and MCF-7 (I–J) cells treated for 8 h with 10 nM E2 or 100 nM G-1 and then switched for additional 8 h to medium without serum in the presence of 10 ng/ml IL1β or conditioned medium collected from CAFs (CM/CAFs) treated for 8 h with vehicle [CM/CAFs (+ vehicle)], 10 nM E2 [CM/CAFs (+ E2)] and 100 nM G-1 [CM/CAFs (+ G-1)]. SkBr3 and MCF-7 cells treated for 8 h with 10 nM E2 or 100 nM G-1 were also exposed to [CM/CAFs (+ E2)] and [CM/CAFs (+ G-1)] alone or in combination with 1 μM IL1R1 antagonist namely IL1R1a. β -actin serves as a loading control. Results shown are representative of at least two independent experiments.

Article Snippet: Equal amounts of whole protein extract were resolved on a 10% SDS-polyacrylamide gel, transferred to a nitrocellulose membrane (Amersham Biosciences, GE Healthcare, Milan, Italy), probed overnight at 4 °C with antibodies against IL1β (R&D Systems, Inc. Celbio, Milan, Italy), IL1R1 (OriGene Technologies, TEMA ricerca srl, Bologna, Italy), GPER (N-15), PGE synthase (S-16) and β -actin (C-2) all purchased from Santa Cruz Biotechnology, DBA, Milan, Italy).

Techniques: Expressing, Control

Journal: Scientific reports

Article Title: GPER signalling in both cancer-associated fibroblasts and breast cancer cells mediates a feedforward IL1β/IL1R1 response.

doi: 10.1038/srep24354

Figure Lengend Snippet: Figure 9. Schematic representation of ligand-activated GPER that generates a feedforward loop coupling IL1β induction by CAFs to IL1R1 expression by cancer cells, toward the induction of IL1β/IL1R1 target genes and biological responses as well as invasive features in breast cancer cells as fibroblastoid cytoarchitecture and F-actin reorganization.

Article Snippet: Equal amounts of whole protein extract were resolved on a 10% SDS-polyacrylamide gel, transferred to a nitrocellulose membrane (Amersham Biosciences, GE Healthcare, Milan, Italy), probed overnight at 4 °C with antibodies against IL1β (R&D Systems, Inc. Celbio, Milan, Italy), IL1R1 (OriGene Technologies, TEMA ricerca srl, Bologna, Italy), GPER (N-15), PGE synthase (S-16) and β -actin (C-2) all purchased from Santa Cruz Biotechnology, DBA, Milan, Italy).

Techniques: Expressing