Journal: bioRxiv

Article Title: Immune-evasion of KRAS -mutant lung adenocarcinoma mediated by cAMP response element-binding protein

doi: 10.1101/2021.06.19.449094

Figure Lengend Snippet: (A-C) Experimental setup (schematics; boxes = months), representative H&E-stained lung sections (images), and pooled results (graphs) from Creb1 WT/WT ( n = 17) and Creb1 F/F ( n = 11) mice (FVB strain) at 6 months after i.t. 5 × 10 9 PFU Ad-CRE followed by i.p. 1 g/Kg urethane 2 weeks later (A), from Scgb1a1.CRE; Creb1 WT/WT ( n = 12) and Scgb1a1.CRE; Creb1 F/F ( n = 9) mice (FVB strain) at 6 months after i.p. 1 g/Kg urethane (B), and from LSL. KRAS G12D ; Creb1 WT/WT ( n = 19) and Scgb1a1.CRE; Creb1 F/F ( n = 19) mice (C57BL/6 strain) at 4 months after i.t. 5 × 10 8 PFU Ad-CRE. Arrows in images denote LUAD. (D) Creb1 F/F LUAD cells were derived from a LUAD induced by four weekly i.p. injections of 1 g/Kg urethane in a FVB Creb1 F/F mouse (schematic; boxes = months), were stably transfected with p LUC or p CRE , were validated, and were assessed for MTT reduction ( n = 4/data-point) and tumor growth in FVB mice (p LUC, n = 12; p CRE, n = 10). (E) HEK293T cells stably transfected with p Kras G12C plus p LUC or p Creb1 , were validated (immunoblots; n, nuclear; c, cytoplasmic) and 3 × 10 6 cells were injected s.c. into NOD/SCID mice( n = 6/group). (D, E) Shown are representative photographs and data summaries as mean (circles) and SD (bars). P , probability, two-way ANOVA. ** and ****: P < 0.01 and P < 0.0001, respectively, compared with p LUC , Bonferroni post-tests. (F, G) Representative May-Grünwald-Giemsa-stained cytocentrifugal specimens (F) and data summary (G; n = 10/group) of bronchoalveolar lavage (BAL) neutrophils (NΦ) from LUAD-bearing Creb1 WT/WT and Creb1 F/F mice. Data in (A-C, G) are shown as raw data points (circles), rotated kernel density plots (violins), medians (dashed lines), and interquartile ranges (dotted lines). P , probability, Mann-Whitney U-test.

Article Snippet: C57BL/6 mouse B16F10 (RRID:CVCL_0159) skin melanoma, PANO2 (RRID:CVCL_D627) pancreatic and Lewis lung carcinomas (LLC, RRID:CVCL_4358), as well as A549 (RRID:CVCL_0023) LUAD cells were from the National Cancer Institute Tumor Repository (Frederick, MD); human HEK293T (RRID:CVCL_0063) embryonic kidney cells were from the American Type Culture Collection (Manassas, VA); C57BL/6 mouse MC38 (RRID:CVCL_B288) colon adenocarcinoma cells were a gift from Dr Barbara Fingleton (Vanderbilt University, Nashville, TN, USA) ( , ).

Techniques: Staining, Derivative Assay, Stable Transfection, Transfection, Western Blot, Injection, MANN-WHITNEY