Fig. 5: Depletion of linc00899 leads to long-lived mitotic microtubules.

a Acetylated α-tubulin levels in HeLa cells after RNAi-mediated linc00899 depletion or treatment with a negative control siRNA (Ctl, from Ambion), based on immunofluorescence after staining with antibodies against α-tubulin (microtubules, red) and acetylated α-tubulin (long-lived microtubules, green). Representative images correspond to maximum intensity projections of confocal micrographs. DNA (blue) was stained with Hoechst. b Acetylated α-tubulin levels after treatment of HeLa cells with 0.05% DMSO or 3 nM taxol (1 h) based on immunofluorescence as in a. Scale bar, 5 µm for a, b. c Quantification of acetylated α-tubulin intensity over the total level of α-tubulin from maximum intensity projections obtained in a, b. Numbers of cells analysed is n = 49 for cells treated with 0.05% DMSO (1 h), n = 30 for cells treated with 0.5 nM taxol (1 h), n = 45 for cells treated with 3 nM taxol (1 h), n = 29 for cells treated with Ctl si and n = 33 for linc00899 RNAi. We also quantified acetylated α-tubulin after 20-h taxol treatment. Numbers of cells analysed is n = 42 for cells treated with 0.05% DMSO (20 h), n = 37 for cells treated with 0.5 nM taxol (20 h), n = 47 for cells treated with 3 nM taxol (20 h), n = 29 for cells treated with Ctl si and n = 32 for linc00899 RNAi. Cells with multipolar spindles were excluded from the analysis. Swarm plots represent values from single mitotic cells with the median denoted by the horizontal line. *P < 0.05, ***P < 0.001 and ****P < 0.0001 by Mann–Whitney test. d EB1 signal in HeLa cells after RNAi-mediated linc00899 depletion or treatment with Ctl si based on immunofluorescence after staining with antibodies against α-tubulin (in red) and EB1 (in green). Images correspond to maximum intensity projections of confocal micrographs. DNA is shown in blue. Scale bar, 10 µm. e Quantification of EB1 intensity from maximum intensity projections obtained in d. Numbers of cells analysed is n = 39 for Ctl si and n = 38 for linc00899 RNAi. Swarm plots represent values from single mitotic cells, with the median represented by the horizontal line. ****P < 0.0001 by Mann–Whitney test. Source data are provided as a Source Data file.