Fig 1: BOD1L1 expression is required for normal kinetochore phosphorylationA) Immunofluorescence images from an asynchronous population of SW620 cells transfected with control shRNA or shRNA against BOD1L1 in prometaphase or metaphase and stained for Hec1, Hec1 pT31 and DAPI. The BOD1L1 and Hec1 channels were adjusted evenly for brightness and contrast for presentation. The DAPI channel of each condition was adjusted independently. Representative images from 2 independent biological experiments are shown. The scale bars of main images are 10 µm, and insets are 1 µm.B) As for panel (A), except for KNL1 pMELT/ACA instead of Hec1 pT31/total Hec1C) Quantification of the relative kinetochore pT31 Hec1/Hec1 intensities from the conditions from panel (A). The condition with the lowest level of pT31 Hec1/Hec1 was set to 1 and the other conditions shown as fold-changes. 25 kinetochores were quantified from each of 20 cells for each of 2 independent biological repeats. Error bars indicate the mean +/- SEM. Statistical significance was calculated between the indicated conditions using Dunnett’s multiple comparison test.D) As for panel (C), except for Hec1 pS44/total Hec1instead of Hec1 pT31/total Hec1E) As for panel (C), except for Hec1 pS55/total Hec1 instead of Hec1 pT31/total Hec1F) As for panel (C), except for DSN1 pS109/ACA instead of Hec1 pT31/total Hec1G) As for panel (C), except for KNL1 pMELT/ACA instead of Hec1 pT31/total Hec1H) As for panel (C), except for MCAK pS95/ACA instead of Hec1 pT31/total Hec1I) As for panel (C), except for Aurora B pT232/ACA instead of Hec1 pT31/total Hec1J) Quantification of total cellular Aurora A pT288/Total Aurora A. The condition with the lowest level of pT31 Hec1/Hec1 was set to 1 and the other conditions shown as fold-changes. The level of protein was measured from at least 25 cells for each of 2 independent biological repeats. Error bars indicate the mean +/- SEM. Statistical significance was calculated between the indicated conditions using Dunnett’s multiple comparison test.