Journal: Cell Death & Disease

Article Title: STING activation induces polarized cytokine secretion of IFN-β and IL-17A promoting photoreceptor death and choroidal disruption in age-related macular degeneration

doi: 10.1038/s41419-026-08491-w

Figure Lengend Snippet: a Schematic of polarized human iPSC-RPE culture in a transwell system. Medium from the insert (upper chamber) was collected as apical media, and media from the bottom chamber as basal media. IFN-β levels in ( b ) apical and ( c ) basal media of iPSC-RPE derived from AMD patients ( n = 3) and their healthy siblings as controls ( n = 3). d Human iPSC-derived three-dimensional retinal organoid at day 180 (D180) of differentiation showing i) structure (brightfield microscopy) and ii) expression of Müller glial marker CRALBP (Green) and photoreceptor markers recoverin (Red), cone arrestin (Green), and rod opsin (Red). e LDH release assay showing the percentage of cytotoxicity in retinal organoids exposed to media derived from iPSC-derived control ( n = 3) and AMD ( n = 3) RPE cultures. f Heatmap showing differential expression of IL-17 pathway genes in RPE from 5- and 10-month-old Cryba1 floxed and Cryba1 cKO mice. IL-17A levels are significantly higher in Cryba1 cKO mice at both time points compared to controls. g H&E staining of retinal sections reveal choroidal and retinal abnormalities in Il17a KI mice. Il17a KI animals differed from WT mice by having marked choroidal thickening with an increased number of melanocytes (lower panel), vacuolated RPE cells (arrows, lower panel), and mild thinning of photoreceptor inner and outer segments ( n = 3). Scale bar= 50 μm (zoomed inset: 20 μm). h Spider plot showing the IS/OS layer thickness across the entire retina Secreted IL-17A levels measured by ELISA in ( i ) basal and ( j ) apical media of polarized iPSC RPE cultures, as well as ( k ) 24-h IFN-β treatment (5000 IU/mL) to iPSC-RPE from AMD subjects ( n = 3) and non-AMD controls ( n = 3). l Secreted IL-17A was measured by ELISA after IFNAR1 inhibition using anifrolumab (100 nM for 24 h) in the apical and basal media of AMD iPSC-derived RPE cells, with no-treatment sibling control iPSC RPE cells used as controls. m western blot analysis was conducted to assess the expression levels of IFNAR1, phospho-STAT1, and phospho-STAT3, along with total STAT1 and STAT3, following IFNAR1 inhibition in AMD iPSC RPE cells. Densitometry analysis ( n = 3 AMD iPSC RPE) quantified the levels of ( n ) phospho-STAT1 and ( o ) phospho-STAT3 upon IFNAR1 inhibition. Values represent mean ± s.d. from triplicate experiments ( n = 3). Abbreviations: REC recoverin, DAPI 4′,6-diamidino-2-phenylindole, CC3 cleaved caspase-3, IS/OS inner/outer segment of photoreceptors, ONL outer nuclear layer, OPL outer plexiform layer, INL inner nuclear layer. Scale bars: 200 μm ( d -i), 20 μm ( d -ii). Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparisons test. ns not significant. *** p < 0.001, **** p < 0.0001.

Article Snippet: Secreted levels of human interferon-beta (IFN-β) and interleukin-17 (IL-17) in the iPSC-RPE conditioned media were measured using Quantikine ELISA kits (R&D Systems, Cat. #DIFNB0 for IFN-β; Cat. #D1700 for IL-17A) according to the manufacturer’s instructions.

Techniques: Derivative Assay, Microscopy, Expressing, Marker, Lactate Dehydrogenase Assay, Control, Quantitative Proteomics, Staining, Enzyme-linked Immunosorbent Assay, Inhibition, Western Blot

Journal: bioRxiv

Article Title: A human arteriovenous differentiation roadmap reveals vein developmental mechanisms and vascular effects of viruses

doi: 10.1101/2025.10.11.681838

Figure Lengend Snippet: Ebola, Nipah, and Andes viruses each exert different effects on hPSC-derived ECs A) Experimental summary. h: hour. B) Replication of Andes, Nipah, and Ebola viruses in hPSC-derived artery and vein ECs, as assayed by qPCR for viral genomes in the culture media. Statistics: unpaired t-test. Error bars: SEM. **P<0.01. n.s.: not significant. †: extensive cell death. C) Bulk RNA-seq of interferon and interferon-stimulated gene expression in hPSC-derived artery and vein ECs 6, 12, 24, and 48 hours after infection with Sendai, Andes, Nipah, and Ebola viruses. Fold change relative to uninfected cells is depicted. D) Bulk RNA-seq of inflammatory cytokine gene expression in hPSC-derived artery and vein ECs 6, 12, 24, and 48 hours after infection with Sendai, Andes, Nipah, and Ebola viruses. Fold change relative to uninfected cells is depicted. E) Summary of the present study. F) Bulk-RNA-seq of hPSC-derived artery ECs 6, 12, 24, and 48 hours after infection with Ebola, Andes, and Sendai viruses, or left uninfected (mock control). Error bars: SEM. G) IFNβ protein secretion by hPSC-derived artery ECs after 24 or 48 hours of infection by Ebola, Andes, and Sendai viruses, or left uninfected (mock control), as measured by ELISA. Statistics: unpaired t-test. **P<0.01. H) Bulk-RNA-seq of hPSC-derived artery ECs 6, 12, 24, and 48 hours after infection with Ebola, Andes, and Sendai viruses, or left uninfected (mock control). Error bars: SEM. Related to Figures S7 and S8.

Article Snippet: Media was then thawed, and the concentration of secreted interferon-β (IFNβ) protein in the 2:1 diluted media was quantified using the Human IFNβ Quantikine QuicKit ELISA Kit (R&D Systems, QK410), as described previously and as per the manufacturer’s instructions.

Techniques: Derivative Assay, RNA Sequencing, Gene Expression, Infection, Control, Enzyme-linked Immunosorbent Assay