Figure 2. HMMR and CHICA are spindle-associated factors required for DYNLL1 targeting. (A) Asynchronous cultures of HeLa cells were fixed and then stained with mouse anti-HMMR, sheep anti-CHICA, rabbit anti-tubulin, and DAPI to detect DNA. Representative examples of the different stages of mitosis are shown. (B) HeLa cells stably expressing GFP-DYNLL1 were transfected with control, CHICA, HMMR, or HURP siRNA duplexes for 72 h, or TACC3 siRNA duplexes for 48 h. The cells were fixed and then stained for CHICA, HMMR, HURP, or TACC3 as appropriate and DAPI to detect DNA. DYNLL1 was visualized using GFP fluorescence. (C) HeLa cells stably expressing GFP-DYNLL1 were transfected with control, DYNLL1, or kinastrin, siRNA duplexes for 72 h, or astrin siRNA duplexes for 48 h. The cells were fixed and then stained for tubulin, CHICA, and DAPI to detect DNA. DYNLL1 was visualized using GFP fluorescence. CHICA-N antibodies were used for all stainings. Bars, 10 µm.
Figure 2.

HMMR and CHICA are spindle-associated factors required for DYNLL1 targeting. (A) Asynchronous cultures of HeLa cells were fixed and then stained with mouse anti-HMMR, sheep anti-CHICA, rabbit anti-tubulin, and DAPI to detect DNA. Representative examples of the different stages of mitosis are shown. (B) HeLa cells stably expressing GFP-DYNLL1 were transfected with control, CHICA, HMMR, or HURP siRNA duplexes for 72 h, or TACC3 siRNA duplexes for 48 h. The cells were fixed and then stained for CHICA, HMMR, HURP, or TACC3 as appropriate and DAPI to detect DNA. DYNLL1 was visualized using GFP fluorescence. (C) HeLa cells stably expressing GFP-DYNLL1 were transfected with control, DYNLL1, or kinastrin, siRNA duplexes for 72 h, or astrin siRNA duplexes for 48 h. The cells were fixed and then stained for tubulin, CHICA, and DAPI to detect DNA. DYNLL1 was visualized using GFP fluorescence. CHICA-N antibodies were used for all stainings. Bars, 10 µm.

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