Journal: PLoS ONE

Article Title: Esrp1 is a marker of mouse fetal germ cells and differentially expressed during spermatogenesis

doi: 10.1371/journal.pone.0190925

Figure Lengend Snippet: Double labelling immunofluorescence experiments with an antibody to Sox9 (A) showed that ESRP1 (B; antibody HPA023719) exhibits minimal labelling of somatic Sertoli cells in the testis. Scale bar: 20 μm all images.

Article Snippet: Antibodies used included two rabbit polyclonal anti-human ESRP1 (HPA023719 and HPA023720; Sigma Aldrich) diluted 1:100 and 1:50 for testis and ovary specimens respectively, a goat polyclonal anti-mouse c-KIT (AF1356, R&D system) diluted 1:250, a goat polyclonal anti-human PLZF (AF2944, R &D system) diluted 1:500, a sheep anti-Sox9 antibody (provided by Dr Dagmar Wilhelm) diluted 1:100 and a mouse monoclonal antibody to the nuclear speckle/spliceosome marker, SC35 (ab11826, AbCam) diluted 1:250.

Techniques: Immunofluorescence

Journal: Oncotarget

Article Title: Loss of zfp36 expression in colorectal cancer correlates to wnt/ β-catenin activity and enhances epithelial-to-mesenchymal transition through upregulation of zeb1, sox9 and macc1

doi: 10.18632/oncotarget.10828

Figure Lengend Snippet: (Panel A ) Boxplot of Log 2 expression values of MACC1, SOX9 and ZEB1 in 23 normal colon mucosa (Normal), 30 primary colon carcinoma (CRC) and 27 liver metastases (Mts) samples. The thick line indicates the median value, the coloured box indicates the interquartile range and the whiskers the minimum and maximum values excluded outliers. Open circles represent data points outside the whiskers. (Panel B ) Schematic representation of the 3′UTRs sequences of MACC1, SOX9 and ZEB1. A-U rich sequences (ARE) are highlighted in bold. (Panel C) HCT116 and SW480 cells were transfected with an empty vector (pCDNA3.1) or a ZFP36-overexpressing vector (pCDNA3.1-ZFP36). RNA was extracted after 48 hours and MACC1, SOX9, ZEB1 mRNA levels were analysed through qRT-PCR analysis. Results are represented as means of three experiments (+/−SEM) and GAPDH was used as endogenous control. * p < 0.05. (Panel D ) HCT116 and SW480 were infected with an empty vector (pRRL) or a ZFP36-overexpressing vector (ZFP36) and corresponding total protein lysates were analysed through Western blotting techniques with antibodies against ZEB1, MACC1, SOX9 and ZFP36. Actin was used as loading control. (Panel E ) A fragment of the 3′UTRs of MACC1, SOX9 and ZEB1 was cloned in a pGL3 vector, downstream of the Luciferase gene. These constructs where co-transfected with a Δ-gal reporter plasmid and with an empty vector (pCDNA3.1) or ZFP36 overexpressing vector (pCDNA3.1-ZFP36) in HEK293T cells. Cells were harvested after 48 hours, luciferase activity was measured and normalized over Δ-gal signals. Results are represented as means of three independent experiments +/−SEM. * p < 0.05, ** p < 0.001, *** p < 0.0001.

Article Snippet: The membranes were then blocked with 5% non-fat milk in TBST 0.1% and immunoblotted overnight at 4°C with different primary antibodies listed below together with their working dilution: N-cadherin (910920 BD Transduction Laboratories); Vimentin (Dako MO725); E-Cadherin (610181 BD); ZO-1 (Zymed 61-7300); MACC1 (Bioss bs-4293R); SOX9 (Bioss bs-4177R); ZEB1 (Sigma HPA027524); ZFP36 (S. Cruz Biotechnology sc-14030).

Techniques: Expressing, Transfection, Plasmid Preparation, Quantitative RT-PCR, Infection, Western Blot, Clone Assay, Luciferase, Construct, Activity Assay

Journal: Cancer Science

Article Title: Frequent PIK3CA mutation in normal endometrial gland drives spheroid formation and may be involved in stem cell propagation

doi: 10.1111/cas.15767

Figure Lengend Snippet: Immunohistochemistry of epithelial stem cell markers and PAX8 in spheroids and endometrium from the same patient. (A) Expression of ALDH1A1, Axin2, and SOX9 in cells composing spheroids. (B) Expression of PAX8 and Axin2 in spheroids and the normal proliferative‐phase endometrium (EM) in the same patient (Case 2). The circle and small square indicate nuclear Axin2‐positive cells at the bottom of the endometrial gland. (C) Merged image with multiple fluorescent‐immunostaining of Axin2, DAPI, and Ki‐67 in the EM of a randomly selected patient. Arrows indicate nuclear (but not cytoplasmic) Axin2‐positive cells at the bottom of the endometrial gland

Article Snippet: After antigen retrieval in sodium citrate buffer, the slides were incubated in overnight at 4°C with Abs at the following dilutions: 1:500 PAX8 (10336‐1‐AP; Proteintech), 1:200 Axin2 (20540‐1‐AP; Proteintech), 1:200 ALDH1A1 (15910‐1‐AP; Proteintech), and 1:500 SOX9 (HPA001758; Atlas Antibodies).

Techniques: Immunohistochemistry, Expressing, Immunostaining

Journal: BMC Veterinary Research

Article Title: Neural, adipocyte and hepatic differentiation potential of primary and secondary hair follicle stem cells isolated from Arbas Cashmere goats

doi: 10.1186/s12917-022-03420-3

Figure Lengend Snippet: Expression of pluripotency-related genes in HFSCs. a Immunofluorescence was used to check the expression of SOX9, OCT4, and SOX2 in PHFSCs; nuclei (blue) and target proteins (green) (Scale bar, 100 µm). b Detection of SOX9, OCT4, and SOX2 expression in SHFSCs by the immunofluorescence (Scale bar, 100 µm). c qRT-PCR was used to evaluate the transcript-level expression of SOX9 , OCT4 , and SOX2 in PHFSCs. PDPCs were used as a control. d qRT-PCR was used to evaluate the transcript-level expression of SOX9 , OCT4 , and SOX2 in SHFSCs. SDPCs were used as a control. e Western blotting was used to check the expression of SOX9, OCT4, and SOX2 in PHFSCs and PDPCs. Full-length blots are presented in Supplementary Fig. . f Western blotting was used to check the expression of SOX9, OCT4, and SOX2 was assessed in SHFSCs and SDPCs. Full-length blots are presented in Supplementary Fig. . g Grey-value quantitative analysis of SOX9, OCT4, and SOX2 expression in PHFSCs. PDPCs were used as a control. h Grey-value quantitative analysis of SOX9, OCT4, and SOX2 expression in SHFSCs. SDPCs were used as a control. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001

Article Snippet: All primary antibodies used in this step and their dilution were as follows: Rabbit Anti-CD34 antibody (Bioss, China) [1:500], Rabbit Anti-CK14 antibody (Bioss, China) [1:1,000], Rabbit Anti-LGR5 antibody (Bioss, China) [1:1,000], Rabbit Anti-K15 antibody (Bioss, China) [1:500], Rabbit Anti-K19 antibody (Bioss, China)[1:1,000], Rabbit Anti-SOX9 antibody (Bioss, China) [1:500], OCT4 Polyclonal Antibody (Proteintech, USA) [1:500], SOX2 Polyclonal Antibody (Proteintech, USA), [1:500], GAPDH Polyclonal Antibody (Proteintech, USA) [1:10000].

Techniques: Expressing, Immunofluorescence, Quantitative RT-PCR, Western Blot

Journal: BMC Veterinary Research

Article Title: Neural, adipocyte and hepatic differentiation potential of primary and secondary hair follicle stem cells isolated from Arbas Cashmere goats

doi: 10.1186/s12917-022-03420-3

Figure Lengend Snippet: The primers of HFSCs markers

Article Snippet: All primary antibodies used in this step and their dilution were as follows: Rabbit Anti-CD34 antibody (Bioss, China) [1:500], Rabbit Anti-CK14 antibody (Bioss, China) [1:1,000], Rabbit Anti-LGR5 antibody (Bioss, China) [1:1,000], Rabbit Anti-K15 antibody (Bioss, China) [1:500], Rabbit Anti-K19 antibody (Bioss, China)[1:1,000], Rabbit Anti-SOX9 antibody (Bioss, China) [1:500], OCT4 Polyclonal Antibody (Proteintech, USA) [1:500], SOX2 Polyclonal Antibody (Proteintech, USA), [1:500], GAPDH Polyclonal Antibody (Proteintech, USA) [1:10000].

Techniques: