Journal: Nature communications

Article Title: IDH1 mutations induce organelle defects via dysregulated phospholipids.

doi: 10.1038/s41467-020-20752-6

Figure Lengend Snippet: Fig. 4 D-2HG levels and SCD expression are responsible for IDH1mut-induced membrane defects in Golgi. a–d Transmission electron micrographs of U251WT, U251R132C U251R132H, and U251R132H + AGI5198 show significant changes in the Golgi structure. Arrows indicate the altered region of Golgi stacks. e Golgi size quantification of U251WT (n = 63, black symbols), U251R132C (n = 59, blue symbols), U251R132H (n = 63, red symbols), and U251R132H + AGI5198 (n = 90, dark red symbols). Statistical values are represented on the graph. f, g Addition of D-2HG in U251WT was enough to cause Golgi dilation. h Golgi size quantification of U251WT (n = 63, black symbols), U251WT + D-2HG (n = 90, green symbols), and U251R132H (n = 63, red symbols). i–j Comparison between Golgi of U251R132H cells and the U251R132H cells that lack hSCD showed a restored Golgi structure. k Inhibiting the hSCD enzyme with CAY10566 led to partial restoration of Golgi structure. l Golgi size quantification of U251R132H (n = 63, red symbols), U251R132H + CAY10566 (n = 69, orange symbols), and siRNA hSCD U251R132H (n = 84, cyan symbols). For e, h, l the mean values and SD are presented and the p values obtained from a one-way ANOVA tests with Tukey’s correction for multiple comparisons were conducted using GraphPad Prism 8.2.1. At least 10 images per organelles were taken. Data are available in Source Data file. Micrographs are representative of three independent experiments.

Article Snippet: Trilencer-27 Fluorescent-labeled transfection control siRNA duplex (OriGENE Inc, SR30002) were used as controls.

Techniques: Expressing, Membrane, Transmission Assay, Comparison

Journal: The Journal of Reproduction and Development

Article Title: Functional roles of NR4A transcription factors in GnRH regulation of gonadotropin gene expression and secretion in rat primary pituitary cells

doi: 10.1262/jrd.2025-100

Figure Lengend Snippet: Effects of Nr4a gene knockdown on gonadotropin gene expression. Primary rat pituitary cells were transfected with siRNAs targeting Nr4a1 (A), Nr4a2 (B), or Nr4a3 (C). Control cells (siCont) were transfected with a non-targeting control siRNA. Gene expression of the gonadotropin subunit was assessed following treatment with or without GnRHa (10 −8 M, three hours). The experiment was repeated at least three times, and representative data are shown. Values are expressed as means ± SEM (n = 4). Statistical significance was determined using the Tukey-Kramer test. * P < 0.05, ** P < 0.01, *** P < 0.001 between siCont and siNr4a1, 2 or 3; ## P < 0.01, ### P < 0.001 vs . Cont.

Article Snippet: Gene-specific 27mer siRNA duplexes targeting Nr4a1 (siNr4a1, #SR500720B), Nr4a2 (siNr4a2, #SR513154C), and Nr4a3 (siNr4a3, #SR500165A) and a universal scrambled negative control siRNA duplex (siCont, #SR30004) were purchased from OriGene Technologies (Rockville, MD, USA).

Techniques: Knockdown, Gene Expression, Transfection, Control

Journal: The Journal of Reproduction and Development

Article Title: Functional roles of NR4A transcription factors in GnRH regulation of gonadotropin gene expression and secretion in rat primary pituitary cells

doi: 10.1262/jrd.2025-100

Figure Lengend Snippet: Effects of Nr4a gene knockdown on gonadotropin secretion. Primary cultured rat pituitary cells were transfected with siRNAs targeting Nr4a1 , Nr4a2 , or Nr4a3 (siCont) and with a control siRNA. After incubation with or without GnRHa (10 −8 M, three hours), FSH (A) and LH (B) concentrations in the culture supernatant were measured. The experiment was repeated at least three times, and representative results are presented. Data are shown as means ± SEM (n = 4). Statistical analysis was conducted using the Tukey-Kramer test. *** P < 0.001 between siCont and siNr4a1, 2 or 3; ### P < 0.001 vs . Cont.

Article Snippet: Gene-specific 27mer siRNA duplexes targeting Nr4a1 (siNr4a1, #SR500720B), Nr4a2 (siNr4a2, #SR513154C), and Nr4a3 (siNr4a3, #SR500165A) and a universal scrambled negative control siRNA duplex (siCont, #SR30004) were purchased from OriGene Technologies (Rockville, MD, USA).

Techniques: Knockdown, Cell Culture, Transfection, Control, Incubation

Journal: bioRxiv

Article Title: NF-κB-Dependent Transcriptional Regulation of Piezo1 Mediates Bacterial Clearance on Stiffened Lung Matrix

doi: 10.1101/2025.11.06.687026

Figure Lengend Snippet: WT, Piezo1 fl/fl , and Piezo1 LysMCre BMDMs were treated with heat-inactivated and live P. aeruginosa (green). Piezo1 Ca 2+ channel activity, phagolysosome maturation, and bacterial clearance were measured on polyacrylamide gels of pathophysiologic range lung stiffness (1 kPa: normal lung, 8-25 kPa: injured lung) and standard tissue culture conditions (10 kPa). ( A ) Phagolysosome maturation was measured using pH-sensitive fluorescent pHrodo bioparticles in P. aeruginosa -treated WT BMDMs on pathophysiologic-range matrix stiffness (1, 8, 25 kPa), n = 5. Phagolysosome maturation is stiffness-dependent and requires matrix stiffness which resembles injured lung (25 kPa). Phagolysosome maturation is reduced upon downregulation of Piezo1 by ( B ) Cre recombinase, as quantified in ( C ), or ( D ) siRNA, as quantified in ( E ), n = 3. ( F ) Bacterial clearance was measured using colony forming units (CFU) of P. aeruginosa . Bacterial clearance is enhanced in Piezo1-sufficient (Piezo1 fl/fl ) BMDMs, n= 3. Data are presented as mean ± SEM. Comparisons by one-way ANOVA or t-test, as appropriate. Scale bars = 25 µm. Images were taken at 40x original magnification. p-values are as indicated.

Article Snippet: BMDMs were given Piezo1-targeted (Origene, Cat. No. SR423525) or control siRNA (Origene, Cat. No. SR30005) and treated with nucleofection solution (Lonza, Cat. No. VCA-1003) prior to nucleofection with program D-032.

Techniques: Activity Assay