Fig 2: Cholesterol depletion hyper-activates LTβR-dependent pro-inflammatory responsea, b The concentrations of secreted CXCL8 were measured with ELISA in media collected from cells preincubated for 1 h with MβCD and then stimulated or not for 4 h (a) or 8 h (b) with Ago or LTα1β2 in the presence or absence of MβCD. Data represent the means ± SEM, n = 4. *P ≤ 0.05; **P ≤ 0.01 by Mann-Whitney or Student’s t-test. c Lysates of A549 cells pretreated for 1 h with MβCD and then stimulated or not for 8 h with LTα1β2 or Ago in the presence or absence of MβCD were analyzed by Western blotting with antibodies against the indicated proteins. Vinculin was used as a loading control. Graph shows densitometric analysis for ICAM1 from Western blotting (protein levels normalized to vinculin). Values are presented as fold change versus controls - unstimulated and untreated cells (black bars). Data represent the means ± SEM, n = 4; ns - P > 0.05; *P ≤ 0.05; **P ≤ 0.01 by one sample t-test (in grey) or by Mann-Whitney (in black). d, f Adhesion of Jurkat, NK cells, neutrophils and T lymphocytes to A549 cells (d) and HUVECs (f) treated as in a or e, respectively. Graphs represent quantification of immune cell adhesion to A549 and HUVECs relative to control (untreated) cells. Values are presented as fold change versus controls - unstimulated and untreated cells (black bars). Data represent the means ± SEM, n = 3 (d), n ≥ 3 (f); ns - P > 0.05; *P ≤ 0.05; **P ≤ 0.01 by one sample t-test. e Lysates of HUVECs preincubated for 1 h with MβCD and then stimulated or not for 6 h with LTα1β2 in the presence or absence of MβCD were analyzed by Western blotting with antibodies against the indicated proteins. Vinculin was used as a loading control.

Fig 3: A CXCL8 expression levels in tumors and control tissues were compared with the GEPIA database. B Visualization of interacting proteins associated with CXCL8. C Interacting protein scores for proteins associated with CXCL8. ACC, adrenocortical carcinoma; BRCA, breast invasive carcinoma; CHOL, cholangiocarcinoma; DLBC, lymphoid neoplasm diffuse large B-cell lymphoma; GBM, glioblastoma multiforme; KICH, kidney chromophobe; KIRP, kidney renal papillary cell carcinoma; LGG, brain lower grade glioma; LUAD, lung adenocarcinoma; OV, ovarian serous cystadenocarcinoma; PCPG, pheochromocytoma and paraganglioma; READ, rectum adenocarcinoma; SKCM, skin cutaneous melanoma; TGCT, testicular germ cell tumors; THYM, thymoma; UCS, uterine carcinosarcoma

Fig 4: Forest plots corresponding to cancer-related risk when assessing the relationship between the CXCL8 -353 polymorphism (AA + AT vs. TT) in all cancers. The squares and horizontal lines respectively correspond to the study-specific ORs and 95% CIs, with square area being indicative of weight (the inverse of the variance). Diamonds additionally reflect the summary OR and 95% CI

Fig 5: CXCL8 expression in tumors and normal tissues for six cancer types (https://ualcan.path.uab.edu/analysis.html). A CXCL8 expression in colon adenocarcinoma (COAD) was elevated relative to normal tissues (P < 0.01). B CXCL8 expression in urothelial carcinoma (URCA) was reduced relative to normal tissues (P < 0.01). C-F CXCL8 expression levels were elevated in tumor tissues as compared to healthy control samples in rectal adenocarcinoma (READ) (C), stomach adenocarcinoma (STAD) (D), thyroid carcinoma (THCA) (E), and head and neck squamous cell carcinoma (HNSC) (F) (All P < 0.01)