Journal: Experimental & Molecular Medicine

Article Title: Reduced expression of TAZ inhibits primary cilium formation in renal glomeruli

doi: 10.1038/s12276-022-00730-2

Figure Lengend Snippet: a Western blots showing IFT88 and IFT140 expression in cells with YAP or TAZ silencing and subjected to 24 h of serum starvation. b Immunocytochemical analysis of IFT140 (red) and the ciliary basal body (green) in SV40MES13 cells with YAP or TAZ knockdown and subjected to 24 h of serum starvation. c The localization of IFT140 in primary cilia was classified into 4 categories: localized only at the ciliary base, localized at both the base and tip, evenly distributed throughout cilia, and not localized in cilia. d NPHP6 and NPHP9 mRNA expression changed in SV40MES13 cells with YAP/TAZ knockdown and subjected to 24 h of serum starvation. e , f Accumulation of NPHP6 and NPHP9 (red) in primary cilia (green) under identical conditions in SV40MES13 cells. g , h The graph shows the fluorescence intensities of NPHP 6 and 9 localized around primary cilia; fluorescence intensities were measured by ZEN blue edition imaging software (Zeiss). i – l Primary cilia recovery in cells with TAZ silencing and knockdown of either NPHP 6 or 9 after 24 h of serum starvation. i Fluorescence images showing primary cilia in cells with knockdown of TAZ or YAP or double knockdown of TAZ and NPHP 6 or 9 after 24 h of serum starvation. j The graphs show the ratio of the number of ciliated cells to the number of DAPI-stained nuclei per image. k Primary cilia lengths were measured by the ZEN black edition program. l The graphs show the proportions of cilia lengths in three ranges: <2.5 μm, 2.5 μm to 5 μm, and >5 μm. a.u. means arbitrary unit. Mean ± SD, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were transfected with siRNAs targeting Nphp6 (sc-72866, Santa Cruz Biotechnology) at a concentration of 30 nM and Nphp9 (50 nM; sc-61177, Santa Cruz Biotechnology) at a concentration of 50 nM; the rest of the transfection process was the same as described above.

Techniques: Western Blot, Expressing, Knockdown, Fluorescence, Imaging, Software, Staining

Journal: Molecular Biology of the Cell

Article Title: The novel centriolar satellite protein SSX2IP targets Cep290 to the ciliary transition zone

doi: 10.1091/mbc.E13-09-0526

Figure Lengend Snippet: Cep290 recruitment to centrosomal satellites and basal bodies depends on SSX2IP in RPE-1 cells. (a) Cells were transfected with control or SSX2IP siRNA and stained for Cep290 and γ-tubulin using indirect immunofluorescence. (b) Average projection of 50 cells immunostained as shown in a. Centering of single images before projection was based on the γ-tubulin signal. (c, d) Quantification of signal intensities of Cep290 at the basal body (c) or in satellites (d) of cells transfected with control or SSX2IP siRNA. (c, d) Left (bars), mean values of averages ± SEM from three independent experiments ( n = 150) normalized to controls. Right (box-and-whiskers plots), quantification of a single representative experiment. * p < 0.05, ** p < 0.01, *** p < 0.001 (two-tailed Student's t test).

Article Snippet: Of importance, mutations in Cep290, also known as BBS14, were previously shown to correlate with several ciliopathies, including Bardet–Biedl syndrome, Meckel–Gruber syndrome, and Joubert syndrome ( Staples et al. , 2012 ).

Techniques: Transfection, Control, Staining, Immunofluorescence, Two Tailed Test