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. 2019 Jul 5;8(7):684. doi: 10.3390/cells8070684

Figure 1.

Figure 1

Spastin localization at midbody is reduced in HIPK2-depleted cells. (A,C) HeLa cells transfected with a mix of three validated HIPK2-specific (siHIPK2) or negative control (siCtr) stealth siRNAs were analyzed by real-time (RT)-PCR, WB and IF 96h post transfection. Molecular weight markers are reported here and in the following figures in kilodalton (kDa). Values are mean ± standard error of mean (SEM) from two independent experiments, each performed in triplicate. ****p < 0.0001, t-test. Representative WB is shown and relative densitometric values of HIPK2/GAPDH are reported below each lane. IF was performed with Abs, recognizing indicated proteins in combination with anti-α- or -β-tubulin, to mark midbody and DAPI to stain DNA. In (B), representative immunostainings of midbody magnification are shown. Bar, 2 µM. In (C), data quantification is shown, values are mean ± SEM from three independent experiments, each performed in duplicate, and >100 midbodies were analyzed per condition. Note that, to avoid alterations of the IF signals, siHIPK2 midbodies showing >2 tubulin-labelled puncta were excluded by this analysis; ***p < 0.001 (p = 0.0002), t-test.