The trichoplein–AurA pathway must suppress primary cilia assembly in order for cells to exit G1.

Photoproduct distribution in films of cinnamate polymers was analyzed to reveal the contribution of both photoisomerization and photodimerization to LC alignment photoregulation. A polymethacrylate with o-cinnamate side chains displayed preferential formation of Z-isomer while the dimerization takes place more favorably for other polymers including poly(vinyl cinnamate). On the basis of the relationship between photoproduct distribution and liquid crystal photoalignment and on the reversibility of the photoinduced reorientation of a liquid crystal, it was concluded that the photoalignment results from the polarization photoisomerization of cinnamate residues in the same manner as that of photochromic moieties like azobenzenes, whereas the (2 + 2) photodimerization plays a role in enhancing the thermal stability of the homogeneous photoalignment.

Supplementary Methods Western blot analysisCells were lysed with Cell Lysis Buffer (Cell Signaling Technologies, Danvers, MA, USA). After cell lysis, lysates were centrifuged at 16,000 g for 5 min at 4°C. The supernatant was used for subsequent procedures. Western blot analyses were conducted after separation by SDS/PAGE electrophoresis and transfer to PVDF membranes. Signal was detected using Chemiluminescence Imaging System (M&S Instruments Inc, Tokyo, Japan). All Western blot experiments were repeated at least twice, and a representative result is shown. RNA-sequencingTotal RNA was extracted from parental and AMG510 resistant NCI-H358 cells using the miRNeasy Mini Kit. An RNAseq library was prepared using the NEBNext ® Ultra™ RNA Library Prep Kit for Illumina ® (New England Biolabs, Ipswich, MA, USA) following the manufacturer's protocols. The enriched libraries were sequenced as 150-bp pairedend reads using NovaSeq (Illumina, San Diego, CA, USA) at Veritas Genetics (Danvers, MA, USA). Transcript-level abundance estimation was conducted using kallisto software (1) with transcript sequences and annotations from GENCODE Release 33 for a pair of RNA-seq fastq files, i.e., Read1 and Read2, for each sample. Next, the transcriptlevel estimates were summarized to gene-level abundance estimates using tximport package (2) on R (3). The matching between transcript IDs (ENST IDs) and gene IDs (ENSG IDs) was based on GENCODE Release 33. Finally, the gene-level abundance estimates of the all samples were merged by tximport and passed to DESeq2 tool (4) to obtain normalized count estimates for all genes of each sample. GSEA analysisGene Set Enrichment Analysis (GSEA) (5) was applied to the obtained normalized count data using GSEA software (Ver. 4.0.3; https://www.gsea-msigdb.org/gsea/index.jsp) with the hallmark and oncogeneic gene set collection in MSigDB 7.1 (6). A gene expression matrix file in gct format was created for each case-control pair from the normalized count data. We used the gene set permutation as the permutation type for this analysis.

Despite the progress in the therapeutic treatment of diabetes mellitus, diabetic nephropathy is one of the major complications of diabetes and the single largest cause of endstage renal diseases. Histologically, early diabetic nephropathy is characterized by glomerular hypertrophy and the inappropriate dilatation of afferent arterioles, which are believed to be associated with hyperfiltration and are followed by thickening of the glomerular basement membrane and accumulation of mesangial matrix [1,2]. The pathogenesis of glomerular hypertrophy and afferent arteriolar dilatation due to diabetes is, however, still not clear.Nitric oxide (NO) has been identified as a pleiotropic intercellular messenger that regulates a variety of cellular functions [3] and an endothelium-derived relaxing factor that inhibits contraction of vascular smooth muscles, as well as the adhesion or aggregation of platelets [4]. Three isoforms of this enzyme have Diabetologia (1998) Summary The overproduction of nitric oxide (NO) is reported in the diabetic kidney and considered to be involved in glomerular hyperfiltration. The precise mechanism of NO production in the diabetic kidney is, however, not known. In this report, we compare the localization of endothelial cell nitric oxide synthase (ecNOS) isoform expression in the kidney tissue of streptozotocin (STZ)-induced diabetic rats and 5/6 nephrectomized rats and clarify the pivotal role of ecNOS for the glomerular hyperfiltration in the early stages of diabetic nephropathy. In diabetic rats, the diameters of afferent arterioles, the glomerular volume, creatinine clearance, and urinary NO 2 / NO 3 were increased after the induction of diabetes. Efferent arterioles were, however, not altered. Insulin or L-NAME treatment returned the diameters of afferent arterioles, glomerular volume, creatinine clearance, and urinary NO 2 /NO 3 to normal. The expression of ecNOS in afferent arterioles and glomeruli of diabetic rats increased during the early stages of the disease, but was not altered in efferent arterioles. Treatment with either insulin or L-NAME decreased ecNOS expression in afferent arterioles and in glomeruli. In contrast, the ecNOS expression was upregulated in both afferent and efferent arterioles and in the glomeruli of 5/6 nephrectomized rats, where the dilatation of afferent and efferent arterioles and glomerular enlargement were observed. Treatment with L-NAME ameliorated the ecNOS expression and dilatation of arterioles. We conclude that enhanced NO synthesis by ecNOS in afferent arterioles and glomerular endothelial cells in response to the hyperglycaemic state could cause preferential dilatation of afferent arterioles, which ultimately induces glomerular enlargement and glomerular hyperfiltration. [Diabetologia (1998 Keywords Nitric oxide (NO), endothelial cell nitric oxide synthase (ecNOS), diabetic nephropathy, afferent arterioles, glomerular hyperfiltration Received: 5 January 1998 and in revised form: 27 April 1998Corresponding author: H. Sugimoto MD., Department of Med...

SummaryThe keratin cytoskeleton performs several functions in epithelial cells and provides regulated interaction sites for scaffold proteins, including trichoplein. Previously, we found that trichoplein was localized on keratin intermediate filaments and desmosomes in welldifferentiated, non-dividing epithelia. Here, we report that trichoplein is widely expressed and has a major function in the correct localization of the centrosomal protein ninein in epithelial and non-epithelial cells. Immunocytochemical analysis also revealed that this protein is concentrated at the subdistal to medial zone of both mother and daughter centrioles. Trichoplein binds the centrosomal proteins Odf2 and ninein, which are localized at the distal to subdistal ends of the mother centriole. Trichoplein depletion abolished the recruitment of ninein, but not Odf2, specifically at the subdistal end. However, Odf2 depletion inhibited the recruitment of trichoplein to a mother centriole, whereas ninein depletion did not. In addition, the depletion of each molecule impaired MT anchoring at the centrosome. These results suggest that trichoplein has a crucial role in MT-anchoring activity at the centrosome in proliferating cells, probably through its complex formation with Odf2 and ninein.