Journal: Medicine

Article Title: Obstetric outcomes of women vaccinated with the COVID-19 vaccine (≥1 dose): A single-center retrospective cohort study of pregnant Chinese women

doi: 10.1097/MD.0000000000039053

Figure Lengend Snippet: Features of studies included in the review.

Article Snippet: Sadarangani et al [ ] , Nov 2021–December 2021 , 8705 (5597 vs 3108) , Prospective , Pregnant females aged 15–49 yr , 3414 Pfizer-BioNTech; 2183 mRNA-1273 , Canada , .

Techniques:

Journal: Cell Biology and Toxicology

Article Title: SP1 and p23 play a crucial role in the circadian target gene induction of activated aryl hydrocarbon receptor in human breast cells

doi: 10.1007/s10565-025-10080-0

Figure Lengend Snippet: Circadian pattern in protein levels of the AHR cofactor SP1. a ) To identify AHR co-factors with a circadian expression level, the mRNA fold change of non-synchronized and synchronized HME1 cells after a treatment with 0.5 nM TCDD was analyzed at the indicated timepoints after synchronization. The mRNA levels of AHR , ARNT , XAP2 , SP1 , HSP90 , P23 and AHRR were analyzed by RT-qPCR. For each timepoint the fold change of the CYP1A1 expression was calculated based on the DMSO control of the respective timepoint. The TCDD mediated induction of AHR , ARNT , XAP2 , SP1 , HSP90 , and P23 remained stable over time in both non-synchronized and synchronized cells. The induction of the AHR target gene AHRR displayed under synchronized conditions a circadian pattern with peaking after 24 h and 48 h. Each bar represents the mean ± SD of three independent experiments. The individual data points from the single experiments are displayed. b) Representative Western blots from three independent experiments of the AHR co-factors AHR, ARNT, XAP2, SP1, HSP90 and p23 from HME1 cells after 12, 24, 36, and 48 h of synchronization. The protein levels of SP1 show a circadian pattern with peaking at 24 h and 48 h upon synchronization indicating that SP1 protein levels under circadian regulation. β-Actin and total protein stain served as loading control. The relative amounts of proteins were derived from the representative Western blot signal intensity and normalized to β-Actin. The quantification was conducted with Image Studio™ Lite Software (LI-COR Biosciences)

Article Snippet: The following primary antibodies diluted in 3% w/v BSA/TBST were used: AHR antibody (1:1000; Abcam; ab190797), ARNT antibody (1:1000; Cell Signaling; #3414), p23 antibody (1:10000; Invitrogen; #MA3-414), HSP90 antibody (1:1000; Cell Signaling; #4877), XAP2 antibody (1:1000; Novus Biologicals; NB100-127), SP1 antibody (1:2000; Novus Biologicals; NB600-233), CLOCK antibody (1:1000; Cell Signaling; #5157), BMAL1 antibody (1:1000; Novus Biologicals; NB100-2288), BMAL1 antibody (1:500; Santa Cruz; sc-365645), β-Actin antibody (1:5000, Abcam, ab8227) and GAPDH antibody (1:1000, Cell signaling; #5174).

Techniques: Expressing, Quantitative RT-PCR, Control, Western Blot, Staining, Derivative Assay, Software

Journal: Cell Biology and Toxicology

Article Title: SP1 and p23 play a crucial role in the circadian target gene induction of activated aryl hydrocarbon receptor in human breast cells

doi: 10.1007/s10565-025-10080-0

Figure Lengend Snippet: Modulator of the circadian regulated AHR activity. a ) Representative fitted curve of the circadian rhythm of M13SV1 cells transiently transfected with a PER2:LUCIFERASE (orange) or BMAL1:LUCIFERASE (blue) circadian reporter plasmid. M13SV1 were transfected via electroporation with the indicated reporter plasmids. 24 h after transfection confluent cells were synchronized by treatment with 1 µM dexamethasone for 1 h and subsequently monitored for 48h. The bioluminescence signal was recorded every 30 min and the obtained measurements was analyzed by the ChronAlyzer software. M13SV1 shows a stable synchronization with an anticyclic expression of the PER2 or BMAL1 control luciferase b) CYP1A1 induction at different time points in synchronized and non-synchronized M13SV1 cells upon treatment with 0.5 nM TCDD. CYP1A1 mRNA after TCDD exposure was analyzed by RT-qPCR at indicated timepoints. Fold change was calculated based on the DMSO control of the respective timepoint. In synchronized cells, the TCDD-mediated CYP1A1 induction displays a circadian pattern peaking at 24 h. Each bar represents the mean ± SD of three independent experiments. The individual data points from the single experiments are displayed. Statistical significance was determined by using an unpaired, one-tailed t-test. c) To determine the knock down efficiency of the targeted genes, the mRNA levels of the indicated genes of interest (GOI) were analyzed by RT-qPCR. The knockdown level was calculated by comparing the GOI with the control siRNA transfected cells. Each bar represents the mean ± SD of three independent experiments. The individual data points from the single experiments are displayed. d) Decrease and increase of CYP1A1 induction was determined in M13SV1 cells transfected with siRNA targeting the AHR or its co-factors ARNT , XAP2 , HSP90 and p23 . The transfected cells were synchronized with 1 µM dexamethasone or left non-synchronized and subsequently exposed to 0.5 nM TCDD for 24 h. CYP1A1 mRNA levels were determined by RT-qPCR and the change in induction was calculated for the siRNA-targeted gene compared to the control siRNA. Each bar represents the mean ± SD of three independent experiments. The individual data points from the single experiments are displayed.

Article Snippet: The following primary antibodies diluted in 3% w/v BSA/TBST were used: AHR antibody (1:1000; Abcam; ab190797), ARNT antibody (1:1000; Cell Signaling; #3414), p23 antibody (1:10000; Invitrogen; #MA3-414), HSP90 antibody (1:1000; Cell Signaling; #4877), XAP2 antibody (1:1000; Novus Biologicals; NB100-127), SP1 antibody (1:2000; Novus Biologicals; NB600-233), CLOCK antibody (1:1000; Cell Signaling; #5157), BMAL1 antibody (1:1000; Novus Biologicals; NB100-2288), BMAL1 antibody (1:500; Santa Cruz; sc-365645), β-Actin antibody (1:5000, Abcam, ab8227) and GAPDH antibody (1:1000, Cell signaling; #5174).

Techniques: Activity Assay, Transfection, Luciferase, Plasmid Preparation, Electroporation, Software, Expressing, Control, Quantitative RT-PCR, One-tailed Test, Knockdown

Journal: Cell Biology and Toxicology

Article Title: SP1 and p23 play a crucial role in the circadian target gene induction of activated aryl hydrocarbon receptor in human breast cells

doi: 10.1007/s10565-025-10080-0

Figure Lengend Snippet: The clock core complex BMAL1/CLOCK directly interacts with the AHR/ARNT complex. HME1 cells were synchronized with 1 µM dexamethasone for 1 h and after 23 h cultivation exposed to 2 nM TCDD for additional 1 h. a) Immunoprecipitation of endogenous AHR. AHR was immunoprecipitated from whole-cell lysates and AHR, ARNT, BMAL1, CLOCK, SP1, p23, HSP90 and GAPDH (as control) were detected on Western blots. Representative Western blots are shown from independent experiments. b) Immunoprecipitation of endogenous BMAL1. BMAL1 was immunoprecipitated from whole-cell lysates and BMAL1, CLOCK, AHR, ARNT, SP1, p23, HSP90 and GAPDH (as control) were detected on Western blots. c) Immunoprecipitation of endogenous CLOCK. CLOCK was immunoprecipitated from whole-cell lysates and CLOCK, BMAL1, AHR, ARNT, SP1, HSP90 and GAPDH (as control) were detected on Western blots. Representative Western blots are shown from independent experiments. For all interaction studies, an antibody mock control was included to determine unspecific bands from the antibody solution.

Article Snippet: The following primary antibodies diluted in 3% w/v BSA/TBST were used: AHR antibody (1:1000; Abcam; ab190797), ARNT antibody (1:1000; Cell Signaling; #3414), p23 antibody (1:10000; Invitrogen; #MA3-414), HSP90 antibody (1:1000; Cell Signaling; #4877), XAP2 antibody (1:1000; Novus Biologicals; NB100-127), SP1 antibody (1:2000; Novus Biologicals; NB600-233), CLOCK antibody (1:1000; Cell Signaling; #5157), BMAL1 antibody (1:1000; Novus Biologicals; NB100-2288), BMAL1 antibody (1:500; Santa Cruz; sc-365645), β-Actin antibody (1:5000, Abcam, ab8227) and GAPDH antibody (1:1000, Cell signaling; #5174).

Techniques: Immunoprecipitation, Control, Western Blot