Journal: Cancers

Article Title: Partial Hepatectomy Promotes the Development of KRASG12V-Induced Hepatocellular Carcinoma in Zebrafish

doi: 10.3390/cancers16101793

Figure Lengend Snippet: The effect of s100a1 knockdown on the development of kras G12V -induced HCC in zebrafish larvae. ( A ) Experiment design of s100a1 knockdown and oncogene induction in Lipan larvae and kras+ larvae. ( B ) Western blot of S100A1with the total protein extracted from MO-injected zebrafish embryos at 24 hpf. β-actin served as a loading control. Each protein sample was pooled from 15 zebrafish embryos. The quantification of the intensity of S100A1normalized by the intensity of β-actin was at the lower panel. The uncropped image of blots is shown in . ( C ) Fluorescence images of morpholino-treated Lipan and kras+ larvae after 48 h of Dox treatment. ( D ) Measurement of liver size based on the fluorescence in ( B ) ( n ≥ 8). Circles, squares, and triangles indicate values of individual samples ( E ) Expression of sox5, sox9a, and sox9b in the kras+ livers at 24 h after PH/sham surgery as determined by RT-qPCR ( n = 3). Scale Bar = 200 μm. ns p > 0.05, * p ≤ 0.05, ** p ≤ 0.01.

Article Snippet: The effect of MO- s100a1 -ATG on S100A1 expression was validated in 1-dpf zebrafish embryos with western blotting using the Rabbit-anti-βactin antibody (bs-0061R, Bioss Antibodies, Woburn, MA, USA) (1:5000) and the Rabbit-anti-S100A1 antibody (50266-RP02; Sino Biological, Beijing, China) (1:1000).

Techniques: Knockdown, Western Blot, Injection, Control, Fluorescence, Expressing, Quantitative RT-PCR

Journal: Scientific reports

Article Title: High androgen level during controlled ovarian stimulation cycle impairs endometrial receptivity in PCOS patients.

doi: 10.1038/s41598-024-74295-7

Figure Lengend Snippet: Fig. 1. Expression of endometrial receptivity marks in infertile women with tubal factor and women with polycystic ovary syndrome (PCOS). (a) Messenger RNA expressions of insulin-like growth factor binding protein 1 (IGFBP-1), epidermal growth factor (EGF), transforming growth factor-beta (TGF β1), leukemia inhibitory factor (LIF), matrix metalloproteinase 9 (MMP9) and integrin β3 in human endometria. Total RNA was extracted from endometrium biopsies of infertile women with tubal factor and PCOS (n = 6 each) and then analyzed by quantitative reverse-transcription polymerase chain reaction. The values are presented as mean ± SEM. **P < .01 and *P < .05 (unpaired t-test). (b,c) IGFBP-1 and LIF protein expression. Tissue lysates (80 mg) were extracted from endometrial tissues of infertile women with tubal factor (control; n = 6) and women with PCOS (PCOS; n = 6), and then subjected to Western blot analysis with the use of anti-LIF, anti-IGFBP-1 or anti-β-actin antibody, as described in the Materials and Methods. Left figure shows one representative experiment among three separate experiments. Right histogram shows relative protein levels of IGFBP-1 and LIF normalized to β-actin (Right). The values are presented as mean ± SEM. *P < .05 (unpaired t-test).

Article Snippet: In brief, the separated samples were transferred to polyvinylidene difluoride membranes and incubated overnight at 4 °C with rabbit polyclonal anti-α-IGFBP-1 (1:1000;13981-1-AP, Proteintech), anti-LIF (1:1000; 26757-1-AP, Proteintech), and human polyclonal anti-βactin (1:0000; 66031-1-Ig Proteintech).

Techniques: Expressing, Binding Assay, Reverse Transcription, Polymerase Chain Reaction, Control, Western Blot

Journal: eLife

Article Title: Genetic variation of putative myokine signaling is dominated by biological sex and sex hormones

doi: 10.7554/eLife.76887

Figure Lengend Snippet: ( A ) Overall study design for integration of gene expression from muscle from 310 humans, single-cell RNA-sequencing (RNA-seq), muscle-specific deletion of Esr1 to infer interorgan coregulatory process across major metabolic tissues. (B–C) Differential expression analysis for sex was performed on all genes corresponding to secreted proteins in skeletal muscle (myokines). The specific genes which showed significant changes in each sex are shown as a volcano plot ( B ) and the relative proportions of myokines corresponding to each category at a least-stringent logistic regression p-value less than 0.05 ( C ). ( D ) For each differential expression category based on sex shown in C, myokines were correlated with all other muscle genes for pathway enrichment. Then the top 10 enriched pathways in males, females, or non-sex specific (by overall significance) were visualized together where number of genes corresponding to each category shown as a relative proportion. ( E ) The same analysis as in D, except instead of myokines being correlated with AR, ESR1 , both hormone receptors, or neither, as compared to correlating with all genes. (F–G) Myokines were binned into two categories based on significant differential expression (logistic regression adjusted p-value < 0.05) between muscle-specific WT and MERKO mice ( F ) or those that showed no change ( G ), then visualized as relative proportions within each category shown in ( C ). ( H ) Midweight bicorrelation (bicor) coefficients (color scheme) and corresponding regression p-values (filled text) are shown for muscle MSTN ~ ESR1 or AR in both sexes (top). Below, correlations are shown for differential expression log2FC (color scheme) and corresponding logistic regression p-values (text fill) for MSTN between sexes in humans or WT vs. MERKO mice. ( I ) Quantification of processed form of myostatin ( , bottom band) relative to β-actin in WT or MERKO muscle in male or female mice. p-Values calculated using a Student’s t-test. (J–K) The top three pathways of genes which significantly (p < 1e-4) correlated with muscle MSTN in males ( J ) or females ( K ). For human data, n = 210 males and n = 100 females. For mouse MERKO vs. WT comparisons, n = 3 mice per group per sex. p-Values from midweight bicorrelations were calculated using the Student’s p-value from WGCNA and logistic regression p-values were calculated using DESeq2. Figure 1—source data 1. Skeletal muscle sex hormone receptor expression between sexes.

Article Snippet: Antibody , Rabbit anti-βactin (Rabbit polyclonal) , Genetex , GTX109639 , (1:1000).

Techniques: Gene Expression, RNA Sequencing, Quantitative Proteomics, Expressing

Journal: eLife

Article Title: Genetic variation of putative myokine signaling is dominated by biological sex and sex hormones

doi: 10.7554/eLife.76887

Figure Lengend Snippet: Full immunoblots shown for skeletal muscle lysate blotted for myostatin (top) or β-actin (bottom) corresponding to different C57BL/6J male (left) or female (right) mice in either WT (floxed) or KO (floxed-cre) for skeletal muscle Esr1 . Band sizes shown to indicate either precursor (top band) or processed/LAP form (bottom band) of myostatin.

Article Snippet: Antibody , Rabbit anti-βactin (Rabbit polyclonal) , Genetex , GTX109639 , (1:1000).

Techniques: Western Blot

Journal: eLife

Article Title: Genetic variation of putative myokine signaling is dominated by biological sex and sex hormones

doi: 10.7554/eLife.76887

Figure Lengend Snippet:

Article Snippet: Antibody , Rabbit anti-βactin (Rabbit polyclonal) , Genetex , GTX109639 , (1:1000).

Techniques:

Journal: Anais da Academia Brasileira de Ciencias

Article Title: Attenuation of oxidative stress, inflammation and apoptosis by ethanolic and aqueous extracts of Crocus sativus L. stigma after chronic constriction injury of rats.

doi: 10.1590/0001-3765201420140067

Figure Lengend Snippet: Figure 2 - The effect of aqueous extract (200 mg/kg) and ethanolic extract (200 mg/kg) of Crocus sativus on the spinal cord protein levels of Bax (21 kDa) and Bcl2 (26 kDa) following western blotting (A) and relative density of Bax/ Bcl2 (B), 3 days after CCI. Administration of extracts was started from the day of surgery and continued for seven consecutive days. The semiquantitative analysis of protein levels was carried out by the “Gel Doc 2000 UV System” (Alliance 4.7). (1): CCI/NS (control); (2): Sham/ saline; (3): CCI /Ethanolic extract (200 mg/kg); (4): CCI/Aqueous extract (200 mg/kg). β-Actin is the loading protein control. Data were mean±SEM (n = 3/group). One-way ANOVA followed by Tukey’s post-hoc test was used for multiple comparisons. *p < 0.05; **P < 0.01: a statistically significant difference when compared to CCI saline-treated rats; ##P < 0.01: Difference between CCI-NS and sham group.

Article Snippet: The probed membranes were then incubated with primary antibodies including: rabbit polyclonal anti-B-cell lymphoma 2 (Bcl2), rabbit polyclonal anti-Bcl-2-associated X protein (Bax) and rabbit polyclonal anti-βactin antibodies (Cell Signaling, 1: 1000) for 1–2 hours at room temperature.

Techniques: Western Blot, Control, Saline