Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: Maternal hypoxia developmentally programs low podocyte endowment in male, but not female offspring.

doi: 10.1002/ar.24369

Figure Lengend Snippet: FIGURE 1 Confocal microscopy of four whole glomeruli at postnatal Day 21 in male normoxic (a) and male hypoxic (b), female normoxic (c), and female hypoxic (d) mice. P57 immunofluorescence (red) indicates podocyte nuclei. Synaptopodin (green) immunostaining indicates podocyte cytoplasm

Article Snippet: One midhilar slice per kidney then underwent 1 hr of antigen retrieval in Dako Citrate Antigen Retrieval Solution (S1699; Dako, Glostrup, Denmark) on a Dako PT Link Instrument (PT10126; Dako) for 60 min at 98 C. Slices were then incubated for 6 days on an orbital shaker (OM8, Ratek, Australia) at 37 C with two primary antibody solutions for podocyte labeling, namely polyclonal rabbit anti-mouse p57 antibody (1:200, Santa Cruz Biotechnology, Santa Cruz, CA; sc-8,298; for labeling of podocyte nuclei) and polyclonal goat anti-mouse synaptopodin (1:400, Santa Cruz Biotechnology, Santa Cruz, CA; sc-21537; for labeling of podocyte cytoplasm).

Techniques: Confocal Microscopy, Immunofluorescence, Immunostaining

Journal: Anatomical record (Hoboken, N.J. : 2007)

Article Title: Maternal hypoxia developmentally programs low podocyte endowment in male, but not female offspring.

doi: 10.1002/ar.24369

Figure Lengend Snippet: FIGURE 2 Complete series of confocal optical sections through a whole glomerulus. (a) P57 immunofluorescence (red) indicates podocyte nuclei. (b) P57 and synaptopodin (green) double immunostaining of podocytes

Article Snippet: One midhilar slice per kidney then underwent 1 hr of antigen retrieval in Dako Citrate Antigen Retrieval Solution (S1699; Dako, Glostrup, Denmark) on a Dako PT Link Instrument (PT10126; Dako) for 60 min at 98 C. Slices were then incubated for 6 days on an orbital shaker (OM8, Ratek, Australia) at 37 C with two primary antibody solutions for podocyte labeling, namely polyclonal rabbit anti-mouse p57 antibody (1:200, Santa Cruz Biotechnology, Santa Cruz, CA; sc-8,298; for labeling of podocyte nuclei) and polyclonal goat anti-mouse synaptopodin (1:400, Santa Cruz Biotechnology, Santa Cruz, CA; sc-21537; for labeling of podocyte cytoplasm).

Techniques: Immunofluorescence, Double Immunostaining

Journal: Nature Communications

Article Title: MYOD-SKP2 axis boosts tumorigenesis in fusion negative rhabdomyosarcoma by preventing differentiation through p57 Kip2 targeting

doi: 10.1038/s41467-023-44130-0

Figure Lengend Snippet: a Representative western blot ( n = 3 independent experiments) of the indicated proteins on RD and JR1 cells infected with either Scrambled (shSCR), SKP2.1 (shSKP2.1) or SKP2.2 (shSKP2.2) lentiviral shRNA at 72 h post-selection. Vinculin is the loading control. b Representative light microscopy pictures of soft agar colony formation assay on RD and JR1 cells treated as in ( a ) and grown for 2 weeks. Scale bar = 50 μm. c Histogram depicts the quantification of soft agar colony numbers per field. n = 3 (RD) and n = 4 (JR1) independent experiments, data presented as mean values ± SD, one-way ANOVA. d Representative light microscopy pictures of single cell colony formation assay on RD and JR1 cells treated as in ( a ) and grown for 2 weeks. Scale bar = 50 μm. e Histogram depicts the quantification of colony numbers per field. n = 3 independent experiments, data presented as mean values ± SD, one-way ANOVA. f Representative light microscopy pictures of β-Galactosidase staining of RD and JR1 cells transfected with either SCR or SKP2 siRNA. Scale bar = 100 μm. g Histogram depicts the quantification of the percentage of senescent cells per field. n = 3 independent experiments, data presented as mean values ± SD, Student’s two-tailed t -test. h Images of JR1 shSCR, shSKP2.1, and shSKP2.2 tumors explanted from mice post euthanasia at 48 days post-inoculation. i Tumor volume of shSCR ( n = 11), shSKP2.1 ( n = 5) and shSKP2.2 ( n = 6) JR1 xenografts assessed by caliper measurement represented in mm 3 followed for 48 days post-inoculation. Data presented as mean values ± SD, two-way ANOVA. j Tumor weight of shSCR ( n = 11), shSKP2.1 ( n = 5) and shSKP2.2 ( n = 6) JR1 xenografts. Box plots show 25th to 75th quartiles, black bar shows the median, and whiskers go down to the smallest value and up to the largest. One-way ANOVA k Representative images ( n = 3 independent experiments) of H&E, p27 Kip1 , p57 Kip2 , MYOG, MyHC, and Ki67 immunohistochemistry of tumor sections from JR1 xenografts expressing either shSCR or shSKP2.2. Scale Bars = 100 μm. Source data are provided as a Source Data file.

Article Snippet: RD and JR1 cells were infected with lentiviral Lenti-ORF clone of CDKN1C (RC209840L3, Origene, Rockville, MD, USA).

Techniques: Western Blot, Infection, shRNA, Selection, Control, Light Microscopy, Soft Agar Assay, Colony Assay, Staining, Transfection, Two Tailed Test, Immunohistochemistry, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Hydatidiform Moles: The Contribution of Ancillary Techniques in Refining Their Histopathological Diagnosis

doi: 10.3390/ijms27010142

Figure Lengend Snippet: p57 immunoexpression. ( a ) p57 immunoexpression in CHM, ×10: negative in villous cytotrophoblast and stromal cells (absent, no maternal genome); ( b ) p57 immunoexpression in PHM, ×10: positive in villous cytotrophoblast and stromal cells (nuclear staining present).

Article Snippet: The following primary antibodies were employed: rabbit polyclonal anti-p57 (clone STJ16100411, dilution 1:300, St John’s Laboratory, London, UK), mouse monoclonal anti-Ki-67 (clone NCL-L-Ki67-MM1, dilution 1:100, Leica Novocastra, Nussloch, Germany), rabbit polyclonal anti-hCG (ab9376, dilution 1:100, Abcam, Cambridge, UK), and mouse monoclonal anti-human E-cadherin (clone NCH-38, ready-to-use, Dako, Glostrup, Denmark).

Techniques: Staining

Journal: Journal of cellular biochemistry

Article Title: MicroRNA-145-5p suppresses cell proliferation, migration, and invasion in upper tract urothelial carcinoma by targeting 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase.

doi: 10.1002/jcb.30449

Figure Lengend Snippet: FIGURE 5 RNA‐sequencing analysis of gene expression by 5‐aminoimidazole‐4‐carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC) knockdown in BFTC909 and KMPC3 cells. (A) miR‐145‐5p overexpression and ATIC knockdown reduced phospho‐ p70 S6KThr389 protein levels in BFTC909 cells. (B) Heatmap showed the top 32 upregulated and downregulated genes in small interfering RNA (siRNA)‐ATIC and siRNA‐control transfected BFTC909 and KPMC3 cells. Log2 expression values were row‐normalized using Z‐scores. Red and green colors indicate decreased and increased expression, respectively. (C) The network of the top 32 putative genes identified by Ingenuity Pathway Analysis (IPA) (D) IPA canonical pathways analyzed the top 32 upregulated and downregulated genes in siRNA‐ATIC and siRNA‐control transfected UTUC cells. Data indicated that these genes were correlated with some diseases, molecular functions, and physiological system functions. (E) ATIC knockdown reduced the protein levels of FN1, Slug, and cyclin A2 and induced p57 in BFTC909 cells (N = 3). (F) Overexpression of miR‐145‐5p decreased the protein levels of FN1, Slug, cyclin A2, and cyclin B1 and increased the expression of p57 in BFTC909 cells (N = 3). Results are shown as the mean ± standard deviation; *p < 0.05 and **p < 0.01. N.S., not significant.

Article Snippet: See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense Technology), fibronectin 1 (FN1) (GTX112794, GeneTex), Slug (NBP2‐52570, Novus), cyclin A2 (#4656, Cell Signaling Technology), cyclin B1 (#4138, Cell Signaling Technology), p57 (NBP1‐89917, Novus), and interferon‐ induced transmembrane 1 (IFITM1) (NBP1‐89345, Novus).

Techniques: RNA Sequencing, Gene Expression, Knockdown, Over Expression, Small Interfering RNA, Control, Transfection, Expressing, Standard Deviation

Journal: Journal of cellular biochemistry

Article Title: MicroRNA-145-5p suppresses cell proliferation, migration, and invasion in upper tract urothelial carcinoma by targeting 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase.

doi: 10.1002/jcb.30449

Figure Lengend Snippet: FIGURE 6 5‐aminoimidazole‐4‐carboxamide ribonucleotide formyltransferase/IMP cyclohydrolase (ATIC) reversed the inhibitory effects of miR‐145‐5p in upper tract urothelial carcinoma (UTUC) cells. (A) miR‐145‐5p inhibited proliferation in BFTC909 and UM‐UC‐14 cells but was restored by ATIC overexpression (N = 3). (B) miR‐145‐5p repressed the migration abilities of BFTC909 and UM‐UC‐14 cells but they were restored by ATIC overexpression (N = 3). (C) miR‐145‐5p suppressed the invasion abilities of BFTC909, UM‐UC‐14, and KMPC3 cells but they were restored by ATIC overexpression (N = 3). (D) Western blot analysis revealed that ATIC, FN1, Slug, cyclin A2, cyclin B1, P57, and IFITM1 levels were restored after the cotransfection of miR‐145‐5p mimics and CMV‐ATIC compared with the levels in BFTC909 cells transfected with miR‐145‐5p alone with α‐tubulin as a reference. (E) Quantification of the protein levels of ATIC, FN1, Slug, cyclin A2, cyclin B1, P57, and IFITM1 from (D) (N = 3). (F) Analysis of miR‐145‐5p, ATIC, IFITM1, CDKN1C, FN1, SNAI2, CCNA2, and CCNB1 expression in UTUC tissues compared with that in adjacent normal tissues using GSE159824 database (N = 2). Data are shown as the mean ± standard deviation; *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: See the T erm s and C onditions (https://onlinelibrary.w iley.com /term s-and-conditions) on W iley O nline L ibrary for rules of use; O A articles are governed by the applicable C reative C om m ons L icense Technology), fibronectin 1 (FN1) (GTX112794, GeneTex), Slug (NBP2‐52570, Novus), cyclin A2 (#4656, Cell Signaling Technology), cyclin B1 (#4138, Cell Signaling Technology), p57 (NBP1‐89917, Novus), and interferon‐ induced transmembrane 1 (IFITM1) (NBP1‐89345, Novus).

Techniques: Over Expression, Migration, Western Blot, Cotransfection, Transfection, Expressing, Standard Deviation