Journal: Indian Journal of Pharmaceutical Sciences

Article Title: Protective Effects of Ginsenoside Rb1 in Rats with Diabetic Cardiomyopathy

doi: 10.36468/pharmaceutical-sciences.967

Figure Lengend Snippet: Fig. 4: Ginsenoside Rb1 attenuated inflammation and suppressed the HB-EGF pathway in DCM rats, (A): RNA expressions of IL6 as determined by quantitative real-time RT-PCR; (B): Protein expressions of HB-EGF in serum as determined by ELISA and (C): Protein expressions of HB-EGF in heart tissue as determined by Western blot Note: Compared to NC, ***p<0.001 and compared to DCM, ###p<0.001

Article Snippet: Human Heparin-Binging Epidermal Growth Factor (HB-EGF) Enzyme-Linked Immunosorbent Assay (ELISA): A Human HB-EGF ELISA kit (Elabscience, China) was used to test for the activity of HB-EGF in plasma, according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Drug delivery

Article Title: In vivo antitumor activity of PHT-427 inhibitor-loaded polymeric nanoparticles in head and neck squamous cell carcinoma.

doi: 10.1080/10717544.2024.2449376

Figure Lengend Snippet: Figure 3. NP-427 downregulates EGFR expression in the HNSCC model. (A) EGFR protein expression was determined by ELISA. (B) Fluorescence micro graphs of representative immunostaining of EGFR-Alexa Fluor546 conjugate in tumor tissue (red) and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI) (blue) (x40), Scale bar 100 µm. (C) EGFR immunofluorescence intensity was ana lyzed by ImageJ software. Experimental groups: control group (CTR) treated with 0.9% saline solution; unloaded nanoparticles group (NP-Ø) with a concen tration of 1 mg/mL; PHT-427 inhibitor-free at concentrations of 0.5 and 1 mg/ mL group (PHT-0.5 and PHT-1, respectively) and loaded nanoparticles with the PHT-427 inhibitor at the concentrations of 0.5 and 1 mg/mL groups (NP-0.5 and NP-1, respectively) for 21 days. The diagrams for EGFR represent the mean ± SD of ng/mL for a and arbitrary units (AU) for B. p < .05 versus CTR and NP-Ø groups. (n = 32).

Article Snippet: Protein extracts were assayed by a different human enzyme-linked immunosorbent assay (elisa) kits by the following proteins: eGFR was quantified using an elisa kit (catalog number: csB-e12124h; cusabio, houston, Usa); akt(ps473) and total akt levels were measured using abcam elisa kit (catalog number: ab126433; abcam, cambridge, UK); Phospho-PDK1 (s241) and total PDK1 by the elisa Kit (catalog number: ab279889) of abcam (cambridge, UK), and human Pi3K by the elisa Kit (catalog number: csB-e08417h, cusabio, houston, Usa) according to the protocol’s instructions. the plates were assessed on the FlUOstar Omega (BMG labtech, Ortenberg, Germany) to 450 nm. these experiments were performed in triplicate for each samples (N = 31) (ctR group (n = 4); NP-Ø group (n = 5); Pht-0.5 group (n = 5); Pht-1 (n = 5); NP-0.5 (n = 6); and NP-1 group (n = 6)).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Fluorescence, Immunostaining, Immunofluorescence, Software, Control, Saline

Journal: Journal of Biological Chemistry

Article Title: The Caenorhabditis elegansmRNA 5′-Capping Enzyme

doi: 10.1074/jbc.m212101200

Figure Lengend Snippet: FIG. 3. The CEL-1 GTase domain can replace Ceg1 in S. cerevi- siae. A, complementation of a double deletion mutant (ceg1cet1) lacking both the GTase and RTPase. The CEG1/CET1 shuffling strain YSB719 (11) was transformed with the following LEU2/2-m plasmids: vector (pAD5); MCE (pGL-MCE, expressing mouse capping enzyme MCE1 from the GPD promoter); CEL-1 (pAD5-CEL1, expressing CEL-1 from the ADH1 promoter); CEL-1-(1–585) (pAD5-CEL-1-(1–585), ex- pressing residues 1–585 of CEL-1 from the ADH1 promoter). Leu

Article Snippet: When overexpressed in ceg1 cet1 strain in combination with MCE-(211–597), cells containing Cet1 formed colonies after 1 day, whereas cells containing either CEL-1-(13–221) or MCE-(1–210) formed colonies after 3 days (Fig. 4A).

Techniques: Mutagenesis, Transformation Assay, Plasmid Preparation, Expressing

Journal: Journal of Biological Chemistry

Article Title: The Caenorhabditis elegansmRNA 5′-Capping Enzyme

doi: 10.1074/jbc.m212101200

Figure Lengend Snippet: FIG. 4. The isolated metazoan RTPase domain can replace Cet1 in S. cerevisiae. A, complementation by plasmid shuffling. YSB719 was transformed with pDB20H-MCE-(211–597). A His isolate was subsequently transformed with the following LEU2/2-m plasmids: vector (pRS425 (81)); CET1 (pAD5-CET1); CEL-1 (13–221) (pAD5-CEL-1-(13–221), expressing CEL-1-(13–221) tagged with an HA epitope from the ADH1 promoter); MCE (1–210) (pAD5-MCE-(1–210), expressing MCE-(1–210) tagged with the HA epitope from the ADH1 promoter). Leu His

Article Snippet: When overexpressed in ceg1 cet1 strain in combination with MCE-(211–597), cells containing Cet1 formed colonies after 1 day, whereas cells containing either CEL-1-(13–221) or MCE-(1–210) formed colonies after 3 days (Fig. 4A).

Techniques: Isolation, Plasmid Preparation, Transformation Assay, Expressing

Journal: Journal of Biological Chemistry

Article Title: The Caenorhabditis elegansmRNA 5′-Capping Enzyme

doi: 10.1074/jbc.m212101200

Figure Lengend Snippet: FIG. 5. The addition of the Ceg1-interacting region from Cet1 onto CEL-1 RTPase domain allows it to function in the presence of Ceg1. YSB719 was transformed with (left) pRSH-CEG1 or (right) pDB20H-MCE-(211–597). His isolates were subsequently transformed with the following LEU2/2 m plasmids: vector (pAD5); CET1 (pAD5- CET1); CET1-(1–225)-CEL-1-(13–221) (pAD5-CET1 (1–225)-CEL-1-(13– 221), expressing a fusion protein containing residues 1–225 of Cet1 and CEL-1-(13–221) tagged with the HA epitope); CET1-(1–265)-CEL-1-(13– 221) (pAD5-CET1 (1–265)-CEL-1-(13–221)); cet1–446-(1–265)-CEL-1-(13– 221) (pAD5-cet1–446-(1–265)-CEL-1-(13–221), expressing a fusion pro- tein consisting of residues 1–265 from the cet1–446 mutant (11) and CEL-1-(13–221) tagged with the HA epitope); and CEL-1-(13–221) (pAD5- CEL-1-(13–221)). Leu His transformants were grown in the presence of 5-FOA, and the plates were incubated for 3 days at 30 °C.

Article Snippet: When overexpressed in ceg1 cet1 strain in combination with MCE-(211–597), cells containing Cet1 formed colonies after 1 day, whereas cells containing either CEL-1-(13–221) or MCE-(1–210) formed colonies after 3 days (Fig. 4A).

Techniques: Transformation Assay, Plasmid Preparation, Expressing, Mutagenesis, Incubation

Journal: Journal of Biological Chemistry

Article Title: The Caenorhabditis elegansmRNA 5′-Capping Enzyme

doi: 10.1074/jbc.m212101200

Figure Lengend Snippet: FIG. 6. RTPase activities of CEL-1-(13–221) mutants. A, purification of recombinant polyhistidine-tagged CEL-1-(13–221) protein. The peak fractions of the heparin-Sepharose CL-6B column were analyzed by SDS-PAGE and visualized by Coomassie Brilliant Blue staining (top panel) or immunoblotting with monoclonal anti-polyhistidine antibody (bottom panel). 400 and 50 ng of protein were loaded for the top and bottom panels, respectively. Lane 1, wild-type CEL-1-(13–221); lane 2, C136S; lane 3, R142K; lane 4, D76N; lane 5, E111Q; lane 6, D112N. For reasons that are not clear, the D76N mutant shows slightly altered mobility. B, RTPase assay. The wild-type and mutated CEL-1-(13–221) proteins were incubated at 30 °C for 10 min with 1 M termini of a [-32P]GTP-labeled 65 nucleotide RNA. Reaction mixtures were analyzed by thin layer chromatography (TLC) on polyethyleneimine-cellulose plates. Released phosphate was detected by autoradiography, and radioactive spots were cut out and quantitated by liquid scintillation counting. Relative amounts of released phosphate were plotted versus protein amount. C, in vivo analysis of CEL-1-(13–221) mutants by plasmid shuffling. YSB719 carrying pDB20H-MCE-(211–597) was transformed with the vector pAD5 or derivatives expressing the indicated CEL-1 alleles. Leu His transformants were tested for growth in the presence of 5-FOA. Plates were incubated for 4 days at 30 °C. D, immunoblot analysis of S. cerevisiae whole-cell extracts. YSB719 cells carrying pDB20H-MCE-(211–597) and CEL-1-(13–221) derivatives were grown in selective media but without shuffling out the CEG1/CET1 plasmid. Extracts were prepared, and 10-g protein was analyzed by SDS-PAGE and immunoblotting with the anti-HA antibody12CA5. Lane 1, pAD5; lane 2, pAD5-CEL-1-(13–221); lane 3, pAD5-CEL- 1-(13–221)C136S; lane 4, pAD5-CEL-1-(13–221)C136A; lane 5, pAD5-CEL-1-(13–221)R142K; lane 6, pAD5-CEL-1-(13–221)R142A; lane 7, pAD5- CEL-1-(13–221)D76N; lane 8, pAD5-CEL-1-(13–221)E111Q; lane 9, pAD5-CEL-1-(13–221)D112N.

Article Snippet: When overexpressed in ceg1 cet1 strain in combination with MCE-(211–597), cells containing Cet1 formed colonies after 1 day, whereas cells containing either CEL-1-(13–221) or MCE-(1–210) formed colonies after 3 days (Fig. 4A).

Techniques: Purification, Recombinant, SDS Page, Staining, Western Blot, Mutagenesis, Incubation, Labeling, Thin Layer Chromatography, Autoradiography, In Vivo, Plasmid Preparation, Transformation Assay, Expressing

Journal: Molecules

Article Title: Revealing Novel Source of Breast Cancer Inhibitors from Seagrass Enhalus acoroides : In Silico and In Vitro Studies

doi: 10.3390/molecules29051082

Figure Lengend Snippet: ΔG of Molecular docking parameter of identified compounds of Seagrass E. acoroides .

Article Snippet: In vitro analysis of HIF-1α, EGFR tyrosine kinase, and HER2 Expressions was carried out in accordance with the manufacturer’s protocol (HIF-1 alpha Monoclonal Antibody (ESEE122), eBioscienceTM; Human EGFR (Epidermal Growth Factor Receptor) ELISA Kit; Elabscience ® for HER2) and established research experimental guidelines [ ].

Techniques: Control

Journal: Molecules

Article Title: Revealing Novel Source of Breast Cancer Inhibitors from Seagrass Enhalus acoroides : In Silico and In Vitro Studies

doi: 10.3390/molecules29051082

Figure Lengend Snippet: Downregulation of HIF-1A, EGFR tyrosine kinase, and HER2 by Seagrass EAE.

Article Snippet: In vitro analysis of HIF-1α, EGFR tyrosine kinase, and HER2 Expressions was carried out in accordance with the manufacturer’s protocol (HIF-1 alpha Monoclonal Antibody (ESEE122), eBioscienceTM; Human EGFR (Epidermal Growth Factor Receptor) ELISA Kit; Elabscience ® for HER2) and established research experimental guidelines [ ].

Techniques:

Journal: Cell reports

Article Title: Granulocyte colony stimulating factor promotes scarless tissue regeneration

doi: 10.1016/j.celrep.2024.114742

Figure Lengend Snippet: (A) Parabiosis between WT:WT ( n = 4, red solid line), CXCR2 KO:CXCR2 KO ( n = 3, blue dotted line), and WT:CXCR2 KO mice ( n = 4, black dotted line). Shown is the percentage of ear hole closure. 2-way ANOVA comparing WT:KO pairs to WT:WT pairs. (B) ELISA measuring cytokine expression in injured WT ( n = 4 for day 3, n = 3 for day 7) and CXCR2 KO ( n = 6 for day 3, n = 3 for day 7) plasma. Unpaired two-tailed Student’s t test. (C) WT ( n = 6), CXCR2 KO ( n = 7) and G-CSF depleted CXCR2 KO ( n = 4) plasma was injected daily into the wound bed of WT mice undergoing WIHN for the first 3 days after injury. Shown are representative photographs and quantification of hair follicles. Scale bars, 100 μM. Unpaired two-tailed Student’s t test. (D) G-CSF ( n = 9) or PBS (control, n = 7 ) was injected daily into the wound bed of WT mice undergoing WIHN for the first 3 days after injury. Representative photographs of whole-mount and scanning electron microscopy demonstrating unpigmented hairs in the center of the healed areas. Right: quantification of hair follicles. Scale bars, 100 μM. Unpaired two-tailed Student’s t test. (E) Representative immunofluorescence images of PBS and G-CSF-injected WT wound beds depicting hair follicle structures (Krt14+, Krt6+) Scale bars, 100 μM. (F) Representative photographs and quantification of scar size of G-CSF-treated ( n = 8) or PBS-treated (control, n = 3 ) stented back wounds at day 28 after injury. Scale bars, 1 mm. Unpaired two-tailed Student’s t test. (G) Representative trichrome-stained tissue sections from G-CSF- or PBS-treated stented back wounds. A black line highlights scar size. (H) Quantification of scar diameter for G-CSF ( n = 6) or PBS-treated ( n = 3) mice. Unpaired two-tailed Student’s t test. (I) Wound fibrosis assessed by picrosirius red staining in G-CSF-treated ( n = 14 sections) or PBS-treated ( n = 10 sections). Unpaired two-tailed Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Mean ± SEM are plotted.

Article Snippet: G-CSF , Med Chem Express , HY-P70608.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Clinical Proteomics, Two Tailed Test, Injection, Control, Electron Microscopy, Immunofluorescence, Staining

Journal: Cell reports

Article Title: Granulocyte colony stimulating factor promotes scarless tissue regeneration

doi: 10.1016/j.celrep.2024.114742

Figure Lengend Snippet: (A) Dot plot demonstrating average expression and percentage of immune cells expressing Csf3r . (B) Analysis of key cell-to-cell interactions between immune cells in the ear skin of WT (salmon color) and CXCR2 KO (blue color) mice. Mac, macrophage; T, T cell. (C) Dot plot demonstrating average gene expression of key genes between WT and CXCR2 KO macrophages. (D) GSEA of macrophage populations in WT and CXCR2 KO wounded skin. (E) Representative images and quantification of immunofluorescence of WT and CXCR2 KO wounded skin for CD80 ( n = 4), COX2 ( n = 4), CD163 ( n = 12 for WT and n = 11 for KO), MRC1 ( n = 10 for WT and n = 6 for KO), and ARG1 ( n = 10 for WT and n = 6 for KO). Cell percentages are calculated with total DAPI+ cells as the denominator. Scale bars, 100 μM. Unpaired two-tailed Student’s t test. (F) Representative H&E immunostaining images and quantification of PBS- and G-CSF-injected stented back wounds of WT mice for CD163 ( n = 7 for PBS and n = 5 for G-CSF), MRC1 ( n = 5 for PBS and n = 7 for G-CSF), CD31 ( n = 8 for PBS and n = 7 for G-CSF) and Ki67 ( n = 5 for PBS and n = 4 for G-CSF). Cell percentages are calculated with total DAPI+ cells as the denominator. Scale bars, 100 μM. Unpaired two-tailed Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.01. Mean ± SEM are plotted.

Article Snippet: G-CSF , Med Chem Express , HY-P70608.

Techniques: Expressing, Gene Expression, Immunofluorescence, Two Tailed Test, Immunostaining, Injection