Figure S2.
Analysis of identified hits. (A) Representative max intensity projection of undeconvolved immunofluorescence images of iKO HeLa cells for control, KIF15, or DLGAP5. Stained for microtubules or DNA. Quantified in Fig. 2 E. Scale bar: 5 µm. (B) Quantification of total tubulin immunofluorescence signal in the KIF15 and DLGAP5 iKO HeLa cells treated with nocodazole (noc). n = 73, 70, 61 across three experiments. (C) Quantification of total spindle tubulin immunofluorescence signal in iKO HeLa cells for control, KIF15 or DLGAP5 treated with paclitaxel (pac). n = 66, 73, 66 across three experiments. (D) Mitotic EB3 speed quantification in control, KIF15 or DLGAP5 iKO HeLa cells expressing td-Tomato EB3. n = 82, 148, 111 kymographs, n = 19, 25, 22 cells across three experiments. (E) Interphase EB3 speed quantification in control, KIF15 or DLGAP5 iKO HeLa cells expressing td-Tomato EB3. n = 82, 148, 111, n = 30, 30, 30 cells across three experiments. (F) Interphase EB3 speed quantification in control, HMMR, or SAMHD1 iKO HeLa cells expressing td-Tomato EB3. n = 86, 171, 142 kymographs, n = 30, 30, 32 cells across three experiments. (G) Mitotic EB3 speed quantification in control, HMMR, or SAMHD1 iKO HeLa cells expressing td-Tomato EB3. n = 82, 110, 130 kymographs, n = 20, 19, 23 cells across three experiments. (H) Representative max intensity projection of undeconvolved immunofluorescence images of iKO HeLa cells for control, HMMR, or SAMHD1. Stained for microtubules or DNA. Quantified in Fig. 3 F. Scale bar: 5 µm. (I) Quantification of total mitotic spindle tubulin immunofluorescence signal in the HMMR and SAMHD1 iKO HeLa cells treated with nocodazole. n = 73, 69, 67 across three experimental replicates. (J) Quantification of total mitotic spindle tubulin immunofluorescence in the HMMR and SAMHD1 iKO HeLa cells treated with paclitaxel. n = 60, 61, 60 across three experimental replicates. (K) Representative confocal image of an interphase HeLa cells showing the localization of GFP-tagged SAMHD1. Scale bar: 5 µm. Statistical tests performed: Welch’s t test (***P = <0.001, ****P = <0.0001). Blue lines indicate the median.

Analysis of identified hits. (A) Representative max intensity projection of undeconvolved immunofluorescence images of iKO HeLa cells for control, KIF15, or DLGAP5. Stained for microtubules or DNA. Quantified in Fig. 2 E. Scale bar: 5 µm. (B) Quantification of total tubulin immunofluorescence signal in the KIF15 and DLGAP5 iKO HeLa cells treated with nocodazole (noc). n = 73, 70, 61 across three experiments. (C) Quantification of total spindle tubulin immunofluorescence signal in iKO HeLa cells for control, KIF15 or DLGAP5 treated with paclitaxel (pac). n = 66, 73, 66 across three experiments. (D) Mitotic EB3 speed quantification in control, KIF15 or DLGAP5 iKO HeLa cells expressing td-Tomato EB3. n = 82, 148, 111 kymographs, n = 19, 25, 22 cells across three experiments. (E) Interphase EB3 speed quantification in control, KIF15 or DLGAP5 iKO HeLa cells expressing td-Tomato EB3. n = 82, 148, 111, n = 30, 30, 30 cells across three experiments. (F) Interphase EB3 speed quantification in control, HMMR, or SAMHD1 iKO HeLa cells expressing td-Tomato EB3. n = 86, 171, 142 kymographs, n = 30, 30, 32 cells across three experiments. (G) Mitotic EB3 speed quantification in control, HMMR, or SAMHD1 iKO HeLa cells expressing td-Tomato EB3. n = 82, 110, 130 kymographs, n = 20, 19, 23 cells across three experiments. (H) Representative max intensity projection of undeconvolved immunofluorescence images of iKO HeLa cells for control, HMMR, or SAMHD1. Stained for microtubules or DNA. Quantified in Fig. 3 F. Scale bar: 5 µm. (I) Quantification of total mitotic spindle tubulin immunofluorescence signal in the HMMR and SAMHD1 iKO HeLa cells treated with nocodazole. n = 73, 69, 67 across three experimental replicates. (J) Quantification of total mitotic spindle tubulin immunofluorescence in the HMMR and SAMHD1 iKO HeLa cells treated with paclitaxel. n = 60, 61, 60 across three experimental replicates. (K) Representative confocal image of an interphase HeLa cells showing the localization of GFP-tagged SAMHD1. Scale bar: 5 µm. Statistical tests performed: Welch’s t test (***P = <0.001, ****P = <0.0001). Blue lines indicate the median.

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