. Throughout the manuscript, these mice are referred to as “Eml4-Alk” mice. Due to the autochthonous nature of the tumor model and a lack of available means to reliably assess lung tumor volume noninvasively, a maximal tumor volume was not directly established for the experiments in this study. Instead, mice were closely monitored to ensure that they did not exhibit signs of morbidity, including weight loss, poor body condition, or labored breathing.
was used to determine pixel intensity-based correlations. Tumors were selected as regions of interest, and thresholds were chosen automatically using the Costes’ method. Co-localization was assessed via the pairwise correlation of pixel intensities within each tumor region of interest.QZ1 was reconstituted to 1 mg/mL in water, then reacted with mPEG-Maleimide, MW 2000 g/mol , for PEG coupling via maleimide-thiol chemistry.
For single cell RNA-seq, CD45+ cell depletion and viability enrichment was performed according to manufacturer’s instructions . For depletion of CD45+ cells, the EasySep™ Mouse CD45 Positive Selection Kit , together with a magnet for holding round-bottom or conical tubes , was used for immunomagnetic positive selection of CD45+ leukocytes from the lung tissue preparation, with the goal of ultimately discarding isolated CD45+ cells.
. At least 100,000 cells from each of the QZ1-hi and QZ1-lo compartments were collected into RPMI-1640 + 2% heat-inactivated FBS and pelleted via centrifugation at 300 ×for 5 min. Cell pellets were lysed in Trizol , and RNA was extracted using RNEasy Mini Kits . Bulk RNA sequencing was performed by the MIT BioMicro Center. Libraries were prepared using the Clontech SMARTer Stranded Total RNAseq Kit , precleaned, and sequenced using an Illumina NextSeq500 on an Illumina NextSeq flow cell.
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