Journal: Cell death discovery

Article Title: Plac8-ERK pathway modulation of monocyte function in sepsis.

doi: 10.1038/s41420-024-02012-4

Figure Lengend Snippet: Fig. 1 Expression of plac8 and ERK activation in peripheral blood mononuclear cells of septic patients and normal individuals. A Flow cytometry analysis of peripheral blood CD14+ and CD16+ cell subsets. B Flow cytometry analysis of CD14+ and CD16+ cell subsets. C qRT- PCR analysis of gene expression. D ELISA analysis of cytokine expression. E Western blot analysis of protein expression. F Western blot analysis of protein expression. G Flow cytometry analysis of cell cycle. H Cell proliferation index PI = (S + G2M)/(G0/1 + S + G2M). * indicates P < 0.05 compared to the normal group. Data is presented as mean ± standard deviation. Data analysis was performed using an independent samples t-test, and the experiment was repeated three times.

Article Snippet: After blocking with 0.5% BSA, the membranes were incubated overnight with primary antibodies: rabbit anti-CD14 (ab182032, 1:1000, Abcam, UK), CD16 (Cat No. 16559-1-AP, 1:1000, Proteintech), plac8 (Cat No. 12284-1-AP, 1:200, Proteintech), ERK1/2 (ab17942, 1:1000, Abcam, UK), p-ERK1/2 (ab223500, 1:1000, Abcam, UK), and GAPDH (ab8245, 1:1000, Abcam, UK).

Techniques: Expressing, Activation Assay, Flow Cytometry, Quantitative RT-PCR, Gene Expression, Enzyme-linked Immunosorbent Assay, Western Blot, Standard Deviation

Journal: Cell death discovery

Article Title: Plac8-ERK pathway modulation of monocyte function in sepsis.

doi: 10.1038/s41420-024-02012-4

Figure Lengend Snippet: Fig. 2 Effects of plac8 on peripheral blood mononuclear cell proliferation in septic patients. A qRT-PCR analysis of gene expression. B Western blot analysis of protein band. C Western blot analysis of protein expression. D ELISA analysis of cytokine expression. E Cell proliferation measured by CCK-8 assay, cell proliferation (%) = [OD (treatment group) −OD (blank group)] / [OD (control group) −OD (blank group)] × 100%. * indicates P < 0.05 compared to si-Plac8-NC or Plac8-NC group. Data is presented as mean ± standard deviation. An independent samples t-test was used to compare two groups, and two-way ANOVA was used to compare at different time points. The experiment was repeated three times.

Article Snippet: After blocking with 0.5% BSA, the membranes were incubated overnight with primary antibodies: rabbit anti-CD14 (ab182032, 1:1000, Abcam, UK), CD16 (Cat No. 16559-1-AP, 1:1000, Proteintech), plac8 (Cat No. 12284-1-AP, 1:200, Proteintech), ERK1/2 (ab17942, 1:1000, Abcam, UK), p-ERK1/2 (ab223500, 1:1000, Abcam, UK), and GAPDH (ab8245, 1:1000, Abcam, UK).

Techniques: Quantitative RT-PCR, Gene Expression, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Control, Standard Deviation

Journal: Cell death discovery

Article Title: Plac8-ERK pathway modulation of monocyte function in sepsis.

doi: 10.1038/s41420-024-02012-4

Figure Lengend Snippet: Fig. 5 Effects of plac8 on monocyte cell survival and proliferation in the mouse model. A Flow cytometry analysis of CD14+ and CD16+ cell subsets. B Flow cytometry analysis of peripheral blood CD14+ and CD16+ cell subsets. C Western blot analysis of protein band. D Western blot analysis of protein expression. E ELISA analysis of cytokine expression. F Detection of cell proliferation. G Flow cytometry analysis of cell cycle. H Cell proliferation index PI. * indicates P < 0.05 compared to si-Plac8-NC or Plac8-NC group. Data is presented as mean ± standard deviation. Independent samples t-test was used to compare two groups, and two-way ANOVA was used for comparison at different time points. The experiment was repeated three times.

Article Snippet: After blocking with 0.5% BSA, the membranes were incubated overnight with primary antibodies: rabbit anti-CD14 (ab182032, 1:1000, Abcam, UK), CD16 (Cat No. 16559-1-AP, 1:1000, Proteintech), plac8 (Cat No. 12284-1-AP, 1:200, Proteintech), ERK1/2 (ab17942, 1:1000, Abcam, UK), p-ERK1/2 (ab223500, 1:1000, Abcam, UK), and GAPDH (ab8245, 1:1000, Abcam, UK).

Techniques: Flow Cytometry, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Standard Deviation, Comparison

Journal: Cell death discovery

Article Title: Plac8-ERK pathway modulation of monocyte function in sepsis.

doi: 10.1038/s41420-024-02012-4

Figure Lengend Snippet: Fig. 6 The role and mechanism of Plac8-mediated ERK signaling pathway activation in the proliferation and activation of peripheral blood monocytes in septic patients.

Article Snippet: After blocking with 0.5% BSA, the membranes were incubated overnight with primary antibodies: rabbit anti-CD14 (ab182032, 1:1000, Abcam, UK), CD16 (Cat No. 16559-1-AP, 1:1000, Proteintech), plac8 (Cat No. 12284-1-AP, 1:200, Proteintech), ERK1/2 (ab17942, 1:1000, Abcam, UK), p-ERK1/2 (ab223500, 1:1000, Abcam, UK), and GAPDH (ab8245, 1:1000, Abcam, UK).

Techniques: Activation Assay

Journal: Cell Death & Disease

Article Title: Effects of short-chain fatty acids in inhibiting HDAC and activating p38 MAPK are critical for promoting B10 cell generation and function

doi: 10.1038/s41419-021-03880-9

Figure Lengend Snippet: A Purified splenic B cells were polarized to Bregs after a 2-day coculture with indicated SCFAs or Trichostatin A (TSA, an HDAC inhibitor with high efficiency) in the presence of LPS; the nuclear protein was prepared and tested for HDAC inhibition by ELISA. B – D B10 cell frequency in purified splenic B cells cultured with or without sodium acetate ( B ), HDAC inhibitors TSA or vorinostat ( C ), or HAT inhibitor (HATi) anacardic acid ( D ) under the existence of CD40 mAb or LPS for 2 days. Results were shown as representative FACS plots or bar graphs. The data are presented as mean ± SD from three independent experiments. * P < 0.05, ** p < 0.01, and *** p < 0.001 compared to Ctrl or as indicated.

Article Snippet: In some experiments, cells were treated with inhibitors or metabolites, including HDACis TSA (1–10 nM; Beyotime) and vorinostat (100–500 nM; MCE), GPR41 agonist AR420626 (1–10 μM; APExBIO), GPR43 agonist (1–10 μM; Sigma-Aldrich) or antagonist GLPG0974 (500 nM; Sigma-Aldrich), GPR109A agonist niacin (1 mM; Sigma-Aldrich), HAT inhibitor anacardic acid (1–10 μM; MCE), ERK1/2 inhibitor U0126 (5 μM; MCE), p38 inhibitor SB 203580 (5 μM; MCE), or JNK inhibitor SP600125 (1 μM; MCE).

Techniques: Purification, Inhibition, Enzyme-linked Immunosorbent Assay, Cell Culture

Journal: Biomolecules

Article Title: Postmortem Stability Analysis of Lipids and Polar Metabolites in Human, Rat, and Mouse Brains

doi: 10.3390/biom15091288

Figure Lengend Snippet: Experiment outline. ( A ). Schematic representation of experimental setup. ( B ). Numbers of detected lipid compounds, grouped according to their biochemical properties. Lipid class abbreviations stand for: AHexCer—acylhexosylceramide; BMP—bis(monoacylglycero)phosphate; CAR—acylcarnitine; CE—cholesteryl ester; Cer—ceramide; CL—cardiolipin; CoQ—coenzyme Q; DG—diacylglycerol; FA—fatty acid; GM—ganglioside; HBMP—hydroxy-bis(monoacylglycero)phosphate; HexCer—hexosylceramide; LPC—lysophosphatidylcholine; LPC-O—lysophosphatidylcholine ether; LPE—lysophosphatidylethanolamine; LPE-O—lysophosphatidylethanolamine ether; LPI—lysophosphatidylinositol; LPS—lysophosphatidylserine; MGDG—monogalactosyldiacylglycerol; NAE—N-acylethanolamine; OxFA—oxidized fatty acid; PC—phosphatidylcholine; PC-O—ether-linked phosphatidylcholine; PE—phosphatidylethanolamine; PE-O—ether-linked phosphatidylethanolamine; PE-P—plasmalogen phosphatidylethanolamine; PG—phosphatidylglycerol; PI—phosphatidylinositol; PMeOH—phosphatidylmethanol; PS—phosphatidylserine; PS-O—ether-linked phosphatidylserine; SHexCer—sulfated hexosylceramide; SM—sphingomyelin; ST—sulfatide; TG—triacylglycerol; TG-O—ether-linked triacylglycerol. A list of the detected lipids is provided in . ( C ). Numbers of detected polar metabolite compounds, grouped according to their biochemical properties. A list of the detected polar metabolites is provided in .

Article Snippet: The extraction buffer (MeOH (Carlo reagents, CAS 67-56-1):MTBE (Sharlab, CAS 1634-04-4), (1:3 ( v / v )) was enriched with internal standards including 0.5 μg/mL triacylglycerol (TAG 15:0/18:1-d7/15:0, Avanti Polar Lipids, 791648C), diacylglycerol (DAG 15:0/18:1-d7, Avanti Polar Lipids, 791647C), ceramide (Cer d18:1-d7/15:0, Avanti Polar Lipids, 860681), lysophosphatidylcholine (LPC 18:1-d7, Avanti Polar Lipids, 791643C), phosphatidylglycerol (PG 15:0/18:1-d7, Avanti Polar Lipids, 791640C), phosphatidylcholine (PC 15:0/18:1-d7, Avanti Polar Lipids, 791637C), phosphatidylethanolamine (PE 15:0/18:1-d7, Avanti Lipids, 791638C), methionine-methyl-13C, d3 (Aldrich 299154), and D-L-Aspartic-acid-4 C13 (Aldrich 488968).

Techniques:

Journal:

Article Title:

doi:

Figure Lengend Snippet: A chemical screen for mitochondrial ceramide-binding proteins yields VDAC1 and -2. a Structure of the photoactive and clickable C 15 -ceramide analog, pacCer. b Mitochondria isolated from HeLa cells were incubated with liposomes containing increasing amounts of pacCer, UV irradiated, and then click reacted with AF647-N 3 . Samples were processed for SDS-PAGE, subjected to in-gel fluorescence (IGF, red), and stained with Coomassie blue (CB, blue). p33•Cer denotes a prominently photolabeled protein band of ~33 kDa. c Strategy for the identification of p33•Cer. d p33•Cer was purified from pacCer-labeled and TAMRA/biotin click-reacted mitochondria using NeutrAvidin-beads, imaged by IGF, excised from the gel, digested by trypsin, and then identified by LC-MS/MS. Data from two independent experiments revealed that p33•Cer corresponds to VDAC1 and VDAC2. e Specificity of anti-VDAC antibodies was validated by immunoblotting of mitochondria isolated from HeLa cells treated with non-silencing (siNS) or VDAC-targeting siRNAs (siVDAC1, siVDAC2). Note that the anti-VDAC3 antibody cross-reacts with VDAC2. f Mitochondria isolated from siVDAC1/2-treated HeLa cells were photolabeled with pacCer, click-reacted with AF647-N 3 , and subjected to IGF analysis followed by CB staining. g Fractions obtained during affinity purification of p33•Cer were subjected to SDS-PAGE, transferred on nitrocellulose, analyzed by on-blot-fluorescence (OBF) and probed with antibodies against VDAC1, TOM40, and p60-Mito. h Fractions obtained during affinity purification of p33•Cer were processed as in g and probed with anti-VDAC2 and anti-VDAC3 antibodies. T total mitochondria extract, FT flow-through, W wash, E eluate

Article Snippet: Next, pacFA was coupled to d - erythro -sphingosine (Enzo Biochem) using a combination of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) and hydroxybenzotriazole (HOBT) as condensing reagents, yielding the photoactivatable and clickable C15-ceramide analog, pacCer (85% overall yield). pacPC was synthesized starting from 1-oleoyl-2-hydroxy- sn -glycero-3-phosphocholine (Avanti Polar Lipids) and pacFA under the action of N , N -dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP) with satisfactory yield (39%). pacDAG was synthesized in three steps starting from 1-oleoyl- sn -glycerol (Santa Cruz Biotechnology).

Techniques: Binding Assay, Isolation, Incubation, Liposomes, Irradiation, SDS Page, Fluorescence, Staining, Purification, Labeling, Liquid Chromatography with Mass Spectroscopy, Western Blot, Affinity Purification

Journal:

Article Title:

doi:

Figure Lengend Snippet: Ceramide binding by VDACs relies on the protonation state of the bilayer-facing Glu. a Space-filling and wireframe models of VDAC1 and VDAC2 simulated with the bilayer-facing Glu residue in a protonated or deprotonated state. Indicated are the volumes for which there is ceramide occupancy greater than 10% (orange) or cholesterol occupancy greater than 20% (yellow). b Distribution of the durations of ceramide contacts with VDAC1 and VDAC2 at the preferred binding site as in a . The y -axis indicates the fraction of the total system time spent in binding events of the duration indicated by x . Summing all points’ y -values yields the fraction of total simulation time when ceramide was bound. c Human VDAC1 and VDAC1 E73Q were produced in E. coli , purified, reconstituted in liposomes, and then photolabeled at the indicated pH with pacCer added from an ethanolic stock. Samples were click-reacted with AF647-N 3 , subjected to SDS-PAGE, and analyzed by IGF and CB staining. d Quantitative analysis of relative pacCer photolabeling efficiencies of reconstituted VDAC1 treated as in c . Data are means ± s.d.; n = 4; ** p < 0.01 by two-tailed paired t -test.

Article Snippet: Next, pacFA was coupled to d - erythro -sphingosine (Enzo Biochem) using a combination of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) and hydroxybenzotriazole (HOBT) as condensing reagents, yielding the photoactivatable and clickable C15-ceramide analog, pacCer (85% overall yield). pacPC was synthesized starting from 1-oleoyl-2-hydroxy- sn -glycero-3-phosphocholine (Avanti Polar Lipids) and pacFA under the action of N , N -dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP) with satisfactory yield (39%). pacDAG was synthesized in three steps starting from 1-oleoyl- sn -glycerol (Santa Cruz Biotechnology).

Techniques: Binding Assay, Residue, Produced, Purification, Liposomes, SDS Page, Staining, Two Tailed Test

Journal:

Article Title:

doi:

Figure Lengend Snippet: The bilayer-facing Glu is a critical determinant of pacCer photolabeling of VDACs. a Human VDAC1, VDAC1 E73Q , VDAC2, and VDAC2 E84Q were produced in E. coli , purified, reconstituted in liposomes, and incubated for 30 min at 37 °C with liposomes containing 1 mol% of pacCer or photoactive and clickable analogs of diacylglycerol (pacDAG), phosphatidylcholine (pacPC), phosphatidylethanolamine (pacPE), or cholesterol (pacChol). Samples were UV irradiated and then click-reacted with AF647-N 3 , subjected to SDS-PAGE, and analyzed by IGF and CB staining. b Quantitative analysis of relative labeling efficiencies of reconstituted VDAC1, VDAC1 E73Q , VDAC2, and VDAC2 E84Q with pacLipids as indicated in a . Data are means ± s.d.; n ≥ 3.

Article Snippet: Next, pacFA was coupled to d - erythro -sphingosine (Enzo Biochem) using a combination of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) and hydroxybenzotriazole (HOBT) as condensing reagents, yielding the photoactivatable and clickable C15-ceramide analog, pacCer (85% overall yield). pacPC was synthesized starting from 1-oleoyl-2-hydroxy- sn -glycero-3-phosphocholine (Avanti Polar Lipids) and pacFA under the action of N , N -dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP) with satisfactory yield (39%). pacDAG was synthesized in three steps starting from 1-oleoyl- sn -glycerol (Santa Cruz Biotechnology).

Techniques: Produced, Purification, Liposomes, Incubation, Irradiation, SDS Page, Staining, Labeling

Journal:

Article Title:

doi:

Figure Lengend Snippet: Competitive inhibition of pacCer photolabeling of VDACs by C 16 -ceramide. a VDAC1, VDAC1 E73Q , VDAC2, and VDAC1 E84Q proteoliposomes were photolabeled with the indicated amount of pacCer or pacChol added from ethanolic stocks. Next, samples were click-reacted with AF647-N 3 , subjected to SDS-PAGE, and analyzed by IGF and CB staining. b VDAC1, VDAC1 E73Q , VDAC2, and VDAC1 E84Q proteoliposomes were photolabeled with 0.2 nmol pacCer or pacChol in the presence of the indicated amounts of natural C 16 -ceramide (Cer) added from ethanolic stocks and then processed as in a . Relative labeling efficiencies were quantified and expressed as % of control (0.2 nmol pacCer or pacChol in the absence of Cer). Data are means ± s.d.; n = 3; * p < 0.05, ** p < 0.01, and *** p < 0.001 by two-tailed paired t -test.

Article Snippet: Next, pacFA was coupled to d - erythro -sphingosine (Enzo Biochem) using a combination of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) and hydroxybenzotriazole (HOBT) as condensing reagents, yielding the photoactivatable and clickable C15-ceramide analog, pacCer (85% overall yield). pacPC was synthesized starting from 1-oleoyl-2-hydroxy- sn -glycero-3-phosphocholine (Avanti Polar Lipids) and pacFA under the action of N , N -dicyclohexylcarbodiimide (DCC) and 4-dimethylaminopyridine (DMAP) with satisfactory yield (39%). pacDAG was synthesized in three steps starting from 1-oleoyl- sn -glycerol (Santa Cruz Biotechnology).

Techniques: Inhibition, SDS Page, Staining, Labeling, Control, Two Tailed Test

Journal: Pharmaceuticals

Article Title: Natural Product-Based Screening for Lead Compounds Targeting SARS CoV-2 M pro

doi: 10.3390/ph16050767

Figure Lengend Snippet: ( a ) Scatter diagram of inhibition rates’ distribution of 2526 compounds to M pro at 80 μM. The gradient inhibition rates of compounds ( b ) fraxetin, ( c ) vanitiolide, ( d ) (−)–gallocatechin gallate, ( e ) hematoxylin, ( f ) wedelolactone, ( g ) β,β−dimethylacrylalkannin, ( h ) hydroxytyrosol ccetate, ( i ) Cholesterol 3–Sulfate Sodium Salt, ( j ) ginkgolic acid C15:1, ( k ) melanin, and ( l ) baicalein to M pro under two conditions; error bars: mean ± S.D. of three independent replicates.

Article Snippet: The Natural Product Library for HTS (catalog number L6000, 50 μL per hole, 10 mM, TargetMol, Shanghai, China, September 2020) was purchased from Shanghai Taosu Biochemical Technology Co., Ltd., including the following: fraxetin (catalog number T2909, TargetMol), vanitiolide (catalog number T0243, TargetMol), (−)–gallocatechin gallate (catalog number T3807, TargetMol), hematoxylin (catalog number T1686, TargetMol), wedelolactone (catalog number T3384, TargetMol), β,β–dimethylacrylalkannin (catalog number T5703, TargetMol), hydroxytyrosol acetate (catalog number T8168, TargetMol), cholesteryl sodium sulfate (catalog number T5239, TargetMol), ginkgolic acid C15:1 (catalog number T6S2123, TargetMol), melanin (catalog number HY-113485, MedChemExpress, Shanghai, China), baicalein (catalog number T2858, TargetMol), sanguinarine (catalog number T2781, TargetMol), corylifol A (catalog number T4S0145, TargetMol), M pro fluorescent substrate (MCA-AVLQSGFR-Lys(Dnp)-Lys-NH2,C223JGC240-1/PE0977, GenScript, Nanjing, China), Sensor Chip SA (10291121, GE Healthcare, Shanghai, China), NHS-PEG12-Biotin (21313, Thermo Scientific, Beijing, China), ultrafiltration centrifuge tube (UFC901096, Millipore), 96-well MicrotiterTM microplate (2305-11, Thermo Scientific), black 96-well plate (655077, Greiner Bio-One, Beijing, China), TritonTM X-100 (T109026, Aladdin, Beijing, China), Eppendorf tube (23-2052LK, Crystalgen, Beijing, China).

Techniques: Inhibition

Journal: Pharmaceuticals

Article Title: Natural Product-Based Screening for Lead Compounds Targeting SARS CoV-2 M pro

doi: 10.3390/ph16050767

Figure Lengend Snippet: SPR-binding dissociation curves of compounds ( a ) hematoxylin (single−cycle), ( b ) (−)–gallocatechin gallate (single–cycle), ( c ) ginkgolic acid C15:1 (multi–cycle), ( d ) fraxetin, ( e ) hydroxytyrosol acetate, ( f ) vanitiolide. The gradient concentrations of the compounds are from low to high: 31.25 μM, 62.5 μM, 125 μM, 250 μM and 500 μM, and the single concentration was measured at 200 μM. The single-cycle kinetics were analyzed using 1:1 binding and the multi-cycle kinetics were analyzed by steady-state affinity.

Article Snippet: The Natural Product Library for HTS (catalog number L6000, 50 μL per hole, 10 mM, TargetMol, Shanghai, China, September 2020) was purchased from Shanghai Taosu Biochemical Technology Co., Ltd., including the following: fraxetin (catalog number T2909, TargetMol), vanitiolide (catalog number T0243, TargetMol), (−)–gallocatechin gallate (catalog number T3807, TargetMol), hematoxylin (catalog number T1686, TargetMol), wedelolactone (catalog number T3384, TargetMol), β,β–dimethylacrylalkannin (catalog number T5703, TargetMol), hydroxytyrosol acetate (catalog number T8168, TargetMol), cholesteryl sodium sulfate (catalog number T5239, TargetMol), ginkgolic acid C15:1 (catalog number T6S2123, TargetMol), melanin (catalog number HY-113485, MedChemExpress, Shanghai, China), baicalein (catalog number T2858, TargetMol), sanguinarine (catalog number T2781, TargetMol), corylifol A (catalog number T4S0145, TargetMol), M pro fluorescent substrate (MCA-AVLQSGFR-Lys(Dnp)-Lys-NH2,C223JGC240-1/PE0977, GenScript, Nanjing, China), Sensor Chip SA (10291121, GE Healthcare, Shanghai, China), NHS-PEG12-Biotin (21313, Thermo Scientific, Beijing, China), ultrafiltration centrifuge tube (UFC901096, Millipore), 96-well MicrotiterTM microplate (2305-11, Thermo Scientific), black 96-well plate (655077, Greiner Bio-One, Beijing, China), TritonTM X-100 (T109026, Aladdin, Beijing, China), Eppendorf tube (23-2052LK, Crystalgen, Beijing, China).

Techniques: Binding Assay, Concentration Assay

Journal: Pharmaceuticals

Article Title: Natural Product-Based Screening for Lead Compounds Targeting SARS CoV-2 M pro

doi: 10.3390/ph16050767

Figure Lengend Snippet: Binding pattern of compounds ( a ) baicalein, ( b ) cholesteryl sodium sulfate, ( c ) ginkgolic acid C15:1, ( d ) wedelolactone, ( e ) (−)–gallocatechin gallate, ( f ) melanin, ( g ) β,β–dimethylacrylalkannin, ( h ) hematoxylin to M pro (PDB code: 7RFS) active site. The red and blue colors on the protein surface are the surface electrostatic potential, with red representing negative charge and blue representing positive charge.

Article Snippet: The Natural Product Library for HTS (catalog number L6000, 50 μL per hole, 10 mM, TargetMol, Shanghai, China, September 2020) was purchased from Shanghai Taosu Biochemical Technology Co., Ltd., including the following: fraxetin (catalog number T2909, TargetMol), vanitiolide (catalog number T0243, TargetMol), (−)–gallocatechin gallate (catalog number T3807, TargetMol), hematoxylin (catalog number T1686, TargetMol), wedelolactone (catalog number T3384, TargetMol), β,β–dimethylacrylalkannin (catalog number T5703, TargetMol), hydroxytyrosol acetate (catalog number T8168, TargetMol), cholesteryl sodium sulfate (catalog number T5239, TargetMol), ginkgolic acid C15:1 (catalog number T6S2123, TargetMol), melanin (catalog number HY-113485, MedChemExpress, Shanghai, China), baicalein (catalog number T2858, TargetMol), sanguinarine (catalog number T2781, TargetMol), corylifol A (catalog number T4S0145, TargetMol), M pro fluorescent substrate (MCA-AVLQSGFR-Lys(Dnp)-Lys-NH2,C223JGC240-1/PE0977, GenScript, Nanjing, China), Sensor Chip SA (10291121, GE Healthcare, Shanghai, China), NHS-PEG12-Biotin (21313, Thermo Scientific, Beijing, China), ultrafiltration centrifuge tube (UFC901096, Millipore), 96-well MicrotiterTM microplate (2305-11, Thermo Scientific), black 96-well plate (655077, Greiner Bio-One, Beijing, China), TritonTM X-100 (T109026, Aladdin, Beijing, China), Eppendorf tube (23-2052LK, Crystalgen, Beijing, China).

Techniques: Binding Assay

Journal: Pharmaceuticals

Article Title: Natural Product-Based Screening for Lead Compounds Targeting SARS CoV-2 M pro

doi: 10.3390/ph16050767

Figure Lengend Snippet: The drug-like properties and pharmacokinetic parameters of screening compounds.

Article Snippet: The Natural Product Library for HTS (catalog number L6000, 50 μL per hole, 10 mM, TargetMol, Shanghai, China, September 2020) was purchased from Shanghai Taosu Biochemical Technology Co., Ltd., including the following: fraxetin (catalog number T2909, TargetMol), vanitiolide (catalog number T0243, TargetMol), (−)–gallocatechin gallate (catalog number T3807, TargetMol), hematoxylin (catalog number T1686, TargetMol), wedelolactone (catalog number T3384, TargetMol), β,β–dimethylacrylalkannin (catalog number T5703, TargetMol), hydroxytyrosol acetate (catalog number T8168, TargetMol), cholesteryl sodium sulfate (catalog number T5239, TargetMol), ginkgolic acid C15:1 (catalog number T6S2123, TargetMol), melanin (catalog number HY-113485, MedChemExpress, Shanghai, China), baicalein (catalog number T2858, TargetMol), sanguinarine (catalog number T2781, TargetMol), corylifol A (catalog number T4S0145, TargetMol), M pro fluorescent substrate (MCA-AVLQSGFR-Lys(Dnp)-Lys-NH2,C223JGC240-1/PE0977, GenScript, Nanjing, China), Sensor Chip SA (10291121, GE Healthcare, Shanghai, China), NHS-PEG12-Biotin (21313, Thermo Scientific, Beijing, China), ultrafiltration centrifuge tube (UFC901096, Millipore), 96-well MicrotiterTM microplate (2305-11, Thermo Scientific), black 96-well plate (655077, Greiner Bio-One, Beijing, China), TritonTM X-100 (T109026, Aladdin, Beijing, China), Eppendorf tube (23-2052LK, Crystalgen, Beijing, China).

Techniques: Molecular Weight, Solubility, Permeability

Journal: Langmuir

Article Title: Contact Killing of Gram-Positive and Gram-Negative Bacteria on PDMS Provided with Immobilized Hyperbranched Antibacterial Coatings

doi: 10.1021/acs.langmuir.9b02549

Figure Lengend Snippet: Figure 1. QAC-coatings on PDMS after crosslinking of the immobilized HBPs with a polyamine (PEI). Curing for 3 h at 125°C yielded a flexible coating (left panel) and for 16 h created a highly crosslinked coating that wrinkles upon bending (right panel).

Article Snippet: TMTC = too many to count. (- means not measured) Sample Bacterial challenge CFU/sample % Reduction 102 103 104 E. coli ATCC 1110 1 Control (PDMS) 63±5 - - 1a HBP-PEI+ 0 6±2 43±4 99.6 E. coli Hu 734 2 Control (PDMS) 73±5 - - 2a HBP-PEI+ 12±7 140±16 TMTC 86.0 2b Control + EDTA 76±8 - - 2c HBP-PEI++EDTA 2±2 12±4 38±8 99.6 E. coli ATCC 15597 3 Control (PDMS) 65±6 - - 3a HBP-PEI+ 6±3 65±8 TMTC 93.5 3b Control + EDTA 76±4 - - 3c HBP-PEI++EDTA 0 3±2 41±5 99.6 P. mirabilis 28 cii 4 Control (PDMS) 83±4 - - 4a HBP-PEI+ 65±5 TMTC TMTC 35 4b Control + EDTA 82±3 - - 4c HBP-PEI++EDTA 17±6 51±7 55±4 99.5 P. mirabilis 296 5 Control (PDMS) 70±5 - - 5a HBP-PEI+ 34±6 110±8 TMTC TMTC 5b Control + EDTA 74±3 - - 5c HBP-PEI++EDTA 6±3 27±5 35±8 99.7 Page 20 of 28 ACS Paragon Plus Environment Langmuir 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 21 Prosthesis.

Techniques:

Journal: Langmuir

Article Title: Contact Killing of Gram-Positive and Gram-Negative Bacteria on PDMS Provided with Immobilized Hyperbranched Antibacterial Coatings

doi: 10.1021/acs.langmuir.9b02549

Figure Lengend Snippet: Figure 2: Phase contrast microscopy images of coated PDMS sheets. Left panel, pristine PDMS, middle panel, after PEI treatment and right panel, PDMS after alkylation of PEI using proton sponge (scale bar is 10µm).

Article Snippet: TMTC = too many to count. (- means not measured) Sample Bacterial challenge CFU/sample % Reduction 102 103 104 E. coli ATCC 1110 1 Control (PDMS) 63±5 - - 1a HBP-PEI+ 0 6±2 43±4 99.6 E. coli Hu 734 2 Control (PDMS) 73±5 - - 2a HBP-PEI+ 12±7 140±16 TMTC 86.0 2b Control + EDTA 76±8 - - 2c HBP-PEI++EDTA 2±2 12±4 38±8 99.6 E. coli ATCC 15597 3 Control (PDMS) 65±6 - - 3a HBP-PEI+ 6±3 65±8 TMTC 93.5 3b Control + EDTA 76±4 - - 3c HBP-PEI++EDTA 0 3±2 41±5 99.6 P. mirabilis 28 cii 4 Control (PDMS) 83±4 - - 4a HBP-PEI+ 65±5 TMTC TMTC 35 4b Control + EDTA 82±3 - - 4c HBP-PEI++EDTA 17±6 51±7 55±4 99.5 P. mirabilis 296 5 Control (PDMS) 70±5 - - 5a HBP-PEI+ 34±6 110±8 TMTC TMTC 5b Control + EDTA 74±3 - - 5c HBP-PEI++EDTA 6±3 27±5 35±8 99.7 Page 20 of 28 ACS Paragon Plus Environment Langmuir 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 21 Prosthesis.

Techniques: Microscopy