Fig. 6

IKKα is required for mutant KRAS-induced malignant pleural effusion. a–c Malignant pleural disease induced by LLC cells (Kras^G12C) stably expressing pNGL NF-κΒ reporter and control shRNA (shC) or shRNA targeting IKKα (shChuk), ΙΚΚβ (shIkbkb), ΙΚΚε (shIkbke), or TBK1 (shTbk1) transcripts (n is given in Table 3). Shown are Kaplan–Meier survival plot (a), data summaries of effusion volume and pleural fluid cells (b), and representative images of effusions (dashed lines) and pleural tumors (t) as well as bioluminescent images at day 13 after pleural injections of the indicated tumor cells (c). d–f Malignant pleural disease induced by MC38 cells (Kras^G13R) stably expressing pNGL NF-κΒ reporter and shC, shChuk, or shIkbkb (n is given in Supplementary Table 3). Shown are immunoblots of cytoplasmic extracts (d), data summaries of effusion volume and pleural fluid cells (e), and representative images of effusions (dashed lines) and pleural tumors (t) as well as representative bioluminescent images at day 13 after pleural injections of the indicated tumor cells (f). g–i Malignant pleural disease induced by PANO2 cells (Kras^WT) stably expressing pNGL NF-κΒ reporter and control plasmid (shC) or plasmid encoding IKKα (pChuk), ΙΚΚβ (pIkbkb), or mutant (pKras^G12C) transcripts (n is given in Supplementary Table 3). Shown are Kaplan–Meier survival plot (g), data summary of effusion volume (h), and representative images of effusions (dashed lines) and pleural tumors (t), hearts (h), and lungs (l), as well as representative bioluminescent images at day 14 after pleural injections of the indicated tumor cells (i). Data are presented as mean ± s.d. P, probability of no difference between cell lines by overall log-rank test (a, g) or one-way ANOVA (b, e, h). ns, single, double, and triple asterisks (*, **, and ***): P > 0.05, P < 0.05, P < 0.01, and P < 0.001, respectively, for the indicated comparisons with control cells by Bonferroni post-tests. Scale bars, 0.5 cm