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: 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