![Figure 1. Loss of LKB1 does not affect invadopodia formation in melanoma cells. (A) Western blot showing lentiviral shRNA knockdown of a nontargeting sequence (NS) or LKB1 (knockdown [KD]) in human A2058 (BRAFV600E/RB null) and mouse GR285 (KrasG12D/p53 null/p16 null) melanoma. (B) Invadopodia formation on Alexa Fluor 647–gelatin. GFP is a marker of knockdown, and cells were stained with Alexa Fluor 568–phalloidin to label actin. (C) Percentage of cells with invadopodia shown as means ± SEM. (A2058: nontargeting sequence, n = 297; KD#1, n = 413; KD#2, n = 300. GR285: nontargeting sequence, n = 321; KD#1, n = 255; KD#2, n = 312.) (D) Live-cell invadopodia assay shows no difference in invadopodia formation over time. Dashed lines show raw data and solid lines are best-fit curves to the data. (Trial #1: nontargeting sequence, n = 60; LKB1 knockdown, n = 60. Trial #2: nontargeting sequence, n = 82, LKB1 knockdown, n = 101.) (E) Invadopodia assay on Alexa Fluor 405–gelatin of WM-266-4 (BRAFV600D/PTEN null) Lifeact (LA)-GFP cells and tdTomato-labeled LKB498 (KrasG12D/Lkb1 null) or LKB1 addback melanoma. Cells were stained with Alexa Fluor 647–phalloidin to label actin. Insets show magnified region of invadopodia. Bars, 20 µm.](https://cdn.rupress.org/rup/content_public/journal/jcb/207/2/10.1083_jcb.201404067/7/jcb_201404067_fig1.jpeg?Expires=1773863143&Signature=Wv90zMMKxvKXomtm~xaQrbC2qspXqjIfg813MVy7AUBC2EvJDPGYxFtVzc-n78S4pcTw9W7CZr1Q1Mc0PNhlMoG8ic-AVik9lV3EnJD~Pa3b8YlLhc4sQC0Z1JYH~czVXiw3mdlaa7y4JPE1JsUQmenOvaGafQxgVmrW0H9feJZ6gv1nt5s~ae1rIwfjc8TsL4oo9vqyg9GjHHkF6QxnHN4rwiFQR0wALERI~VbemRbBfMrcobhYqqBl1-XwHTFlUX~sWxl8woYZZN99z3lnovzzhD01PuxUZk79Z4IAQdXqdPlImP4Nd5YcT6MxYlfMphZBDE9ng7v4ez986WmMtQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Loss of LKB1 does not affect invadopodia formation in melanoma cells. (A) Western blot showing lentiviral shRNA knockdown of a nontargeting sequence (NS) or LKB1 (knockdown [KD]) in human A2058 (BRAFV600E/RB null) and mouse GR285 (KrasG12D/p53 null/p16 null) melanoma. (B) Invadopodia formation on Alexa Fluor 647–gelatin. GFP is a marker of knockdown, and cells were stained with Alexa Fluor 568–phalloidin to label actin. (C) Percentage of cells with invadopodia shown as means ± SEM. (A2058: nontargeting sequence, n = 297; KD#1, n = 413; KD#2, n = 300. GR285: nontargeting sequence, n = 321; KD#1, n = 255; KD#2, n = 312.) (D) Live-cell invadopodia assay shows no difference in invadopodia formation over time. Dashed lines show raw data and solid lines are best-fit curves to the data. (Trial #1: nontargeting sequence, n = 60; LKB1 knockdown, n = 60. Trial #2: nontargeting sequence, n = 82, LKB1 knockdown, n = 101.) (E) Invadopodia assay on Alexa Fluor 405–gelatin of WM-266-4 (BRAFV600D/PTEN null) Lifeact (LA)-GFP cells and tdTomato-labeled LKB498 (KrasG12D/Lkb1 null) or LKB1 addback melanoma. Cells were stained with Alexa Fluor 647–phalloidin to label actin. Insets show magnified region of invadopodia. Bars, 20 µm.
