After treatment, cells were harvested and total RNA was extracted via the RNeasy Mini Kit (Qiagen, Valencia, CA) and reverse transcribed with a High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems). The TaqMan Gene Expression kit (Life Technologies) and StepOne Plus (Applied Biosystems) were employed for quantitative analyses. Taqman primer probes for ACTA2 (Hs00426835_g1), SERPINE1 (Hs00167155_m1), COL1A1 (Hs00164004_m1), FN1 (Hs01549976_m1), CCN2 (Hs1026927_g1), EDN1 (Hs00174961_m1), FSTL1 (Hs00907496_m1), TSC22D3 (Hs00608272_m1), NR3C1 (Hs00353740_m1), YAP1 (Hs00902712_g1), and GAPDH (Hs02758991_g1) were employed (all from Thermo Fisher Scientific, Waltham, MA). The ΔΔCt method was employed with GAPDH as the housekeeping gene for quantification of relative expression.
Fstl1
Manufactured by Thermo Fisher Scientific
FSTL1 is a lab equipment product offered by Thermo Fisher Scientific. It is a protein that functions as a regulator of cellular processes. The core function of FSTL1 is to modulate various biological activities, but a detailed description of its intended use is not available.
Automatically generated - may contain errors
Lab products found in correlation
Taqman gene expression kit, by Thermo Fisher Scientific (1 mentions)
Nr3c1, by Thermo Fisher Scientific (1 mentions)
High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (1 mentions)
Gapdh, by Thermo Fisher Scientific (1 mentions)
Steponeplus, by Thermo Fisher Scientific (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Activin a, by Thermo Fisher Scientific (1 mentions)
Mbs1131960, by MyBioSource (1 mentions)
Nb100 1597, by Novus Biologicals (1 mentions)
Ta343590, by OriGene (1 mentions)
2 protocols using fstl1
1
Quantitative Gene Expression Analysis
2
Implantation Protein Profiling in BBT-EVs
Check if the same lab product or an alternative is used in the 5 most similar protocols
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To assess the changes in implantation protein profiles in BBT-EVs as a result of BBT-MSC in vitro communication, pre-confluent 100 mm2 tissue culture dishes of BBT-9 and BBT-18 were cultured for 48 h in media supplemented with MSC-EVs isolated by SEC from stimulated or unstimulated MSC. Confluent 100 mm2 tissue culture dishes of eMSC or pbMSC were cultured for 48 h with or without IFN-τ stimulation (MBS1131960, Mybiosource) (1000 ng/mL). Then, EVs from the different eMSC or pbMSC lines were pooled separately. In addition, to assess uterine cytokines effect on BBT-EVs, pre-confluent 100 mm2 tissue culture dishes of BBT-9 and BBT-18 were cultured in media supplemented with Activin A (PHC9564, Thermo Fisher Scientific) (100 ng/mL), Activin A, and FSTL1 (10924H08H50, Thermo Fisher Scientific) (100 and 50 ng/mL, respectively) or without cytokines during 48 h. The experimental design is shown in Figure 5 . Bead-assisted flow cytometry was performed on the BBT-EVs obtained following the procedures described to analyze the implantation marker expression of MMP2 (MA1-772, Thermo Fisher Scientific); HSPH1 (PA5-77793, Thermo Fisher Scientific); TDGF1 (NB100-1597, Novus Biologicals), and PEG3 (TA343590, Acris-antibodies).
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