Journal: JCI Insight
Article Title: EGFR -mutant transformed small cell lung cancer harbors intratumoral heterogeneity targetable with MEK inhibitor combination therapy
doi: 10.1172/jci.insight.197008
Figure Lengend Snippet: ( A ) Heatmap of cell viability after 5 days’ treatment with 1 μmol/L of each single agent (rows) per various lung cancer cell lines (columns) relative to DMSO control. Commercially available agents used are in . ( B ) Bar graph of IC 50 values of various indicated lung cancer cell lines in response to trametinib. IC 50 of NE and non-NE tSCLC cell lines is shown in red and blue, respectively. ( C ) Dose-response curve of trametinib in tSCLC cell lines (blue: NE, red: non-NE). Cell viability was assessed by CTG after 120 hours ( n = 6, technical replicate, mean ± SD). ( D ) NE tSCLC cell viability was assayed by CTG following treatment with indicated concentration of LIF and IL-6 for 5 days. ( E ) qPCR analysis of LIF mRNA expression levels in non-NE tSCLC cell lines treated with 10 nmol/L trametinib 2 and 6 hours. Gene expression was normalized to GUSB and shown as relative to that of DMSO-treated control ( n = 3, technical replicate, mean ± SD). ****= P ≤ 0.001 by 2-way ANOVA with Šídák’s multiple comparisons test. ( F ) Various cell lines were analyzed for LIF expression by ELISA. Data are presented as mean ± SD ( n = 3. Technical replicates). ( G ) GFP-expressing DFCI112F and DFCI190F cells were incubated with CM from their corresponding adherent cell line pairs pretreated with either LIF neutralizing antibody (500 ng/mL) or control IgG and monitored in real time via IncuCyte imager for green fluorescence intensity. ( H and I ) GFP expressing DFCI112F ( H ) and DFCI190 ( I ) cells were incubated with CM from their adherent cell line pairs pretreated with siRNA against LIF or unrelated sequence and monitored via IncuCyte imager. LIF knockdown efficiency was tested using ELISA ( n = 3, technical replicate, mean ± SD). ( J ) The schema illustrating the interaction between NE and non-NE tSCLC cells through LIF.
Article Snippet: LIF neutralizing antibody (AF-250-NA), IL-6 neutralizing antibody (AF-206-NA), and control antibody (AB-108-C) were purchased from R&D Systems.
Techniques: Control, Concentration Assay, Expressing, Gene Expression, Enzyme-linked Immunosorbent Assay, Incubation, Fluorescence, Sequencing, Knockdown
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