Journal: International Journal of Molecular Sciences

Article Title: Lgr5 Does Not Vary Throughout the Menstrual Cycle in Endometriotic Human Eutopic Endometrium

doi: 10.3390/ijms20010022

Figure Lengend Snippet: Immunofluorescence measurement of LGR5 expression throughout the menstrual cycle in eutopic endometrium. ( A ) Epithelial and stromal expression of LGR5 in control and endometriosis tissue in five phases of the menstrual cycle. ( B ) Total tissue expression across five phases of the menstrual cycle in control and endometriosis tissues. ( C ) Differences in LGR5 expression throughout the menstrual cycle between control and endometriosis tissue groups ( t -test for each phase; proliferative: p = 0.0242; secretory: p = 0.0424; menstruation: p = 0.0121). (Control: n = 24; endometriosis: n = 24). ( D ) Example of immunofluorescence of LGR5 in eutopic endometrium (early secretory phase). In blue: DAPI; in red: LGR5. * p < 0.05.

Article Snippet: Samples stained with monoclonal rabbit anti-LGR5 primary antibody (1 μL per million cells was used; BioNova Scientific, Fremont, CA, USA) and goat Alexa647 anti-rabbit secondary antibody (Invitrogen) at a 1:500 dilution.

Techniques: Immunofluorescence, Expressing, Control

Journal: International Journal of Molecular Sciences

Article Title: Lgr5 Does Not Vary Throughout the Menstrual Cycle in Endometriotic Human Eutopic Endometrium

doi: 10.3390/ijms20010022

Figure Lengend Snippet: In vitro analysis of LGR5 expression throughout the menstrual cycle in endometrial stromal fibroblast primary culture after treatment (E 2 and E 2 P 4 for six and six more days, respectively). ( A ) Left panels show expression of CYR61 (marker of the proliferative phase) and DKK1 (marker of the secretory phase) in the control group ( n = 4); right panels show expression of CYR61 and DKK1 in the endometriosis group ( n = 3). ( B ) Upper panel shows expression of LGR5 in proliferative (E 2 ) and secretory (E 2 P 4 ) phases of the control group; lower panel shows LGR5 expression in both phases of the endometriosis group (E 2 : estradiol; P 4 : progesterone).

Article Snippet: Samples stained with monoclonal rabbit anti-LGR5 primary antibody (1 μL per million cells was used; BioNova Scientific, Fremont, CA, USA) and goat Alexa647 anti-rabbit secondary antibody (Invitrogen) at a 1:500 dilution.

Techniques: In Vitro, Expressing, Marker, Control

Journal: International Journal of Molecular Sciences

Article Title: Lgr5 Does Not Vary Throughout the Menstrual Cycle in Endometriotic Human Eutopic Endometrium

doi: 10.3390/ijms20010022

Figure Lengend Snippet: Percentage of FACS-sorted LGR5 + cells. ( A ) Percentages of LGR5 + cells in different phases of the cycle (proliferative: n = 9; secretory: n = 4; ovulatory: n = 12; ND: n = 9). ( B ) Percentage of LGR5 + cells in eutopic endometrium of different types of endometriosis and controls (ovarian: n = 8; pelvic: n = 3; adenomyosis: n = 4; DIE: n = 10; control: n = 12). ( C ) Percentage of LGR5 + cells identified in eutopic endometrium of women with and without endometriosis (control: n = 12; endometriosis: n = 25). ( D ) Differences in LGR5 + cells between eutopic and ectopic endometrium. Left panel: ovarian endometriosis (eutopic: n = 8; ectopic: n = 4; p = 0.0286). Right panel: DIE (eutopic: n = 10; ectopic: n = 6; p = 0.0411). ( E ) Difference between ovarian and DIE ectopic endometrium (ovarian: n = 4; DIE: n = 6; p = 0.0381). ND: non-determined; DIE: deep infiltrating endometriosis. * p < 0.05, ** p < 0.03.

Article Snippet: Samples stained with monoclonal rabbit anti-LGR5 primary antibody (1 μL per million cells was used; BioNova Scientific, Fremont, CA, USA) and goat Alexa647 anti-rabbit secondary antibody (Invitrogen) at a 1:500 dilution.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Lgr5 Does Not Vary Throughout the Menstrual Cycle in Endometriotic Human Eutopic Endometrium

doi: 10.3390/ijms20010022

Figure Lengend Snippet: Primers used for RT-qPCR.

Article Snippet: Samples stained with monoclonal rabbit anti-LGR5 primary antibody (1 μL per million cells was used; BioNova Scientific, Fremont, CA, USA) and goat Alexa647 anti-rabbit secondary antibody (Invitrogen) at a 1:500 dilution.

Techniques: Sequencing

Journal: PLoS Genetics

Article Title: Whole exome sequencing reveals HSPA1L as a genetic risk factor for spontaneous preterm birth

doi: 10.1371/journal.pgen.1007394

Figure Lengend Snippet: Most significant pathway results for Finnish mothers with recurrent SPTB (n = 10).

Article Snippet: Membranes were incubated overnight in +4°C with primary monoclonal rabbit antibodies against HSPA1L (dilution 1:1000; GTX111045; GeneTex), GR (dilution 1:1000; #3660S; Cell Signaling Technologies) or, as control, β-actin (dilution 1: 20,000; A5060; Sigma) in 5% nonfat dry milk TBS-0.1%Tween.

Techniques:

Journal: PLoS Genetics

Article Title: Whole exome sequencing reveals HSPA1L as a genetic risk factor for spontaneous preterm birth

doi: 10.1371/journal.pgen.1007394

Figure Lengend Snippet: Pathway results for Danish sister pairs (n = 93); showing only genes related to the Discovery findings.

Article Snippet: Membranes were incubated overnight in +4°C with primary monoclonal rabbit antibodies against HSPA1L (dilution 1:1000; GTX111045; GeneTex), GR (dilution 1:1000; #3660S; Cell Signaling Technologies) or, as control, β-actin (dilution 1: 20,000; A5060; Sigma) in 5% nonfat dry milk TBS-0.1%Tween.

Techniques:

Journal: PLoS Genetics

Article Title: Whole exome sequencing reveals HSPA1L as a genetic risk factor for spontaneous preterm birth

doi: 10.1371/journal.pgen.1007394

Figure Lengend Snippet: HSPA1L variants in the large GWAS data and their association to SPTB.

Article Snippet: Membranes were incubated overnight in +4°C with primary monoclonal rabbit antibodies against HSPA1L (dilution 1:1000; GTX111045; GeneTex), GR (dilution 1:1000; #3660S; Cell Signaling Technologies) or, as control, β-actin (dilution 1: 20,000; A5060; Sigma) in 5% nonfat dry milk TBS-0.1%Tween.

Techniques:

Journal: PLoS Genetics

Article Title: Whole exome sequencing reveals HSPA1L as a genetic risk factor for spontaneous preterm birth

doi: 10.1371/journal.pgen.1007394

Figure Lengend Snippet: Functional predictions for rare missense HSPA1L variants.

Article Snippet: Membranes were incubated overnight in +4°C with primary monoclonal rabbit antibodies against HSPA1L (dilution 1:1000; GTX111045; GeneTex), GR (dilution 1:1000; #3660S; Cell Signaling Technologies) or, as control, β-actin (dilution 1: 20,000; A5060; Sigma) in 5% nonfat dry milk TBS-0.1%Tween.

Techniques: Functional Assay, Variant Assay, In Silico

Journal: PLoS Genetics

Article Title: Whole exome sequencing reveals HSPA1L as a genetic risk factor for spontaneous preterm birth

doi: 10.1371/journal.pgen.1007394

Figure Lengend Snippet: (A) HSPA1L is a 641-amino acid protein that consist of two major functional domains; an N-terminal nucleotide-binding domain and a C-terminal substrate-binding domain, that are connected with interdomain linker. Locations of four HSPA1L variants from whole exome sequencing (Discovery and Replication findings) are shown in the protein sequence. Purple diamonds represent ATP nucleotide-binding sites at positions 14−17, 204−206, 270−277 and 341−344. Green circles denote the additional phosphorylation site generated by Ala268Thr and the existing one T267-p. (B) In silico comparison of higher order assembly of reference and modified (Ala268Thr) HSPA1L protein models containing an ADP molecule. Overlayed reference and Ala268Thr molecules are presented as gold and light blue rounded ribbon structures, respectively. The interacting ADP molecule is shown as a stick model. (C) Closeup view of the intermolecular contact interface of HSPA1L bound to the ADP molecule. Key interacting residues (amino acids) are shown and their corresponding side chains are presented as stick molecules; nitrogen and oxygen atoms are indicated in blue and red, respectively. All interacting residues; THR16, TYR17, GLU270, LYS273, ARG274, SER277 (not shown in figure) and ASP368, that bind to the ADP ligand showed small changes in the chemical bond lengths. Only changes of ≥0.002Å are shown in the figure (+/- lenght of Ala268Thr structure in relation to reference structure).

Article Snippet: Membranes were incubated overnight in +4°C with primary monoclonal rabbit antibodies against HSPA1L (dilution 1:1000; GTX111045; GeneTex), GR (dilution 1:1000; #3660S; Cell Signaling Technologies) or, as control, β-actin (dilution 1: 20,000; A5060; Sigma) in 5% nonfat dry milk TBS-0.1%Tween.

Techniques: Functional Assay, Binding Assay, Sequencing, Phospho-proteomics, Generated, In Silico, Comparison, Modification

Journal: PLoS Genetics

Article Title: Whole exome sequencing reveals HSPA1L as a genetic risk factor for spontaneous preterm birth

doi: 10.1371/journal.pgen.1007394

Figure Lengend Snippet: Cultured ESFs were transfected with WT or Ala268Thr HSPA1L -pcDNA3.1 constructs or with empty pcDNA3.1 vector (control). Cells were treated with decidualization media supplemented with 100nM dexamethasone (glucocorticoids) for 72h. Both cytosolic and nuclear protein were extracted, and HSPA1L and GR protein levels were measured by Western blot. Band intensity of HSPA1L or GR was normalized to band intensity of the corresponding β-actin. Cytosolic (A) and nuclear (B) HSPA1L levels as well as cytosolic (C) and nuclear (D) GR levels are shown for control (empty vector), WT and Ala268Thr sample groups. Each experiment was performed as triplicates in three different passages (n = 9 each group, except n = 8 for nuclear control group) and bars represent mean + SEM. Significant p-value <0.05 is presented with an asterisk. Cytosolic GR levels were significantly higher (p = 0.04) in the WT compared to the Ala268Thr group as well as in the WT compared to the control group (p = 0.04).

Article Snippet: Membranes were incubated overnight in +4°C with primary monoclonal rabbit antibodies against HSPA1L (dilution 1:1000; GTX111045; GeneTex), GR (dilution 1:1000; #3660S; Cell Signaling Technologies) or, as control, β-actin (dilution 1: 20,000; A5060; Sigma) in 5% nonfat dry milk TBS-0.1%Tween.

Techniques: Cell Culture, Transfection, Construct, Plasmid Preparation, Control, Western Blot

Journal: Investigative Ophthalmology & Visual Science

Article Title: Differential Expression of Sex-Steroid Receptors in the Choroid Aligns With Central Serous Chorioretinopathy Sex Prevalence Across Different Ages

doi: 10.1167/iovs.65.8.5

Figure Lengend Snippet: List of Antibodies for the Detection of Sex-Steroid Receptors

Article Snippet: Anti-AR (detecting AR ) , Primary , Clone SP107, rabbit monoclonal primary antibody , #760–4605 , OptiView DAB detection kit .

Techniques: