Fig 1: Metastatic cell growth and lung colonization do not require KMT2 complex components.(A) Schematic of subunit composition of several KMT2 complexes. (B) Western blot analyses of the indicated proteins in LM2 cells transduced with inducible shRNA targeting KMT2A, RBBP5, DPY30, HCFC1, CXXC1, and WDR82. Cells were collected after 3 days of doxycycline (DOX) (1 µg/mL) treatment. (C) RT-quantitative PCR (qPCR) analysis of BOD1L1 in LM2 cells transduced with two independent hairpins targeting BOD1L1. Cells were collected after 3 days of DOX treatment (n=4, unpaired two-side Student’s t test). (D–E) Colony formation assay of LM2 shCtrl or shKMT2A cells (shKMT2A-1 and shKMT2A-2) after 9 days of treatment with control or DOX. Representative images (D) and quantification (E) are shown (n=3, unpaired two-side Student’s t test). (F) Normalized bioluminescence unit (BLU) signals of lung metastasis at day 66 of mice injected intravenously with LM2 cells from (B) and fed with DOX chow. The data represent mean ± SEM. Significance determined using unpaired two-tailed Mann–Whitney test. ns, not significant; p=0.2857 (shCtrl versus shKMT2A-1) and p=0.5556 (shCtrl versus shKMT2A-2). (G) RT-qPCR analysis of the noted RP genes in LM2 cells transduced with the indicated inducible shRNAs. Cells were collected after 3 days of DOX treatment. Significance determined by comparing each treatment to shCtrl. (H–I) Colony formation assay of LM2 cells from (G) after 9 days of either control or DOX treatment. Representative images (H) and quantification (I) are shown (n=3, unpaired two-side Student’s t test). ***p=0.0005 (shHCFC1 #1 -DOX versus shHCFC1 #1 +DOX) and **p=0.0039 (shHCFC1 #2 -DOX versus shHCFC1 #2 +DOX). For gel source data, see Figure 5—source data 1. Figure 5—source data 1.Original western blots for Figure 5B.