Extended Data Fig. 9: WSTF in acute and chronic inflammation.

a, IMR90 were treated with TNFα (20 ng/mL) and THP-1 were treated with LPS (1 μg/mL) for 24 h. b, IMR90 treated with IFNβ were analysed by RT-qPCR. Mean values with s.d.; n = 3 biologically independent samples; exact P values are shown; n.s.: non-significant. c-d, cGAS knockout BJ fibroblasts were chronically transfected by ISD (5 μg/mL) (c); IMR90 with MAVS knockdown were chronically transfected by poly I:C (0.25 μg/mL). e-f, BiFC assay in acute and chronic inflammation. g, mCherry-GFP-WSTF ARPE19 were transfected with poly I:C (0.25 μg/mL) chronically. h, IMR90 with FIP200 knockdown were transfected by ISD chronically. i-k, Co-IP in IMR90. l, ATMi rescued WSTF loss in IMR90 under chronic poly I:C or ISD transfection. m, p-ATM in IMR90. n, Sequences of synthesized peptides. o-p, GST-GABARAP pulldown of in vitro translated WSTF with 20 μg/mL competing peptides. q, PHYRE2 prediction of the hydrophobic helix of WSTF aligned with Cav2.2. r, Alphafold2 prediction of WSTF. GABARAP-binding region is highlighted, forming a hydrophobic helix with high confidence (top). ClusPro modelling of GABARAP UDS binding to WSTF helix (bottom). s, UDS and LDS regions on GABARAP and LC3B. t, Co-IP of IMR90 whole cell extracts treated with NLS-cell penetrating peptides in cultured media for 48 h. u, Fractionation control for Fig. 3i. v-w, IMR90 undergoing chronic poly I:C or ISD transfection were treated with NLS-CPP-peptides. Mean values with s.d.; n = 3 biologically independent samples; exact P values are shown; n.s.: non-significant. x, NLS-CPP-WSTF rescued WSTF in senescent IMR90. y, Sequences of mouse peptides. z-aa, Ms NLS-CPP-WSTF rescued WSTF in senescent AML12 and reduced the SASP gene expression. P values were calculated using one-way ANOVA coupled with Dunnett’s post hoc test. The experiments shown in a, c, d-m, o, p, t, u, x, z were repeated independently at least three times with similar results.