Journal: bioRxiv

Article Title: Lipids Maintain Genomic Stability and Developmental Potency of Murine Pluripotent Stem Cells

doi: 10.1101/2022.08.12.503780

Figure Lengend Snippet: a-b, Western blotting for Erk1/2 and p-Erk1/2 proteins in ESCs cultured in 2iL or 2iLA at passage 3 (a) or for Mek/Erk signaling related proteins in ESCs after switching 2iL to 2iLA (b). m, minutes; h: hours. c, qRT-PCR for the naive marker genes Nanog , Prdm14 and formative marker genes Dnmt3b , Pim2 for WT , Erk2 -/- , Erk2-Res ESCs at passage 4 cultured in 2iL or 2iLA. Erk2-Res : Erk2 -rescued in the Erk1/2 mutant ESCs (Lentiviral-based shRNA against Erk1 in the Erk2 -/- ESCs). n= 3 experiments. ns, no significant difference, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, student t-test. d-e, Schematic illustration of the experimental design and ESC colony morphology change (d), and qRT-PCR for the naive marker genes Zfp42 , Prdm14 , and the formative marker genes Dnmt3b , Pim2 for WT, and Erk2 -/- Erk2-Res (e) after 2i and LIF removal for 72 hours. n= 3 experiments. f-g, FACS analysis of X G X T reporter female ESCs cultured in SL2i, 2iL or 2iLA from P1 to over P15 (f) or in 2iL, 2iLA, 2iLA + VX-11e (Erk2 inhibitor) or CLA + VX-11e from P1 to P15 (g), CLA: CHIR 99021 +LIF+ AlbuMAX. h, Karyotyping of the X G X T , Erk2-/- , Mek1 -/- Mek2 -/- double knockout ( Mek1/2 dKO) PSCs cultured in 2iLA over passage 15. n=∼20 metaphases for each cell line.

Article Snippet: The following primary antibodies were used at the indicated dilutions: Zscan4 (Millipore, AB4340); Dnmt3a (Abcam, ab2850), Dnmt3b (Abcam, ab2851), GAPDH (CST, 5174, 1:10,000); Erk1/2 (CST, 4695); p-Erk1/2 (CST, 9101); Nanog (CST, D2A3 XP®).

Techniques: Western Blot, Cell Culture, Quantitative RT-PCR, Marker, Mutagenesis, shRNA, Double Knockout

Journal: bioRxiv

Article Title: Lipids Maintain Genomic Stability and Developmental Potency of Murine Pluripotent Stem Cells

doi: 10.1101/2022.08.12.503780

Figure Lengend Snippet: a, qRT-PCR for Dnmts transcript levels in ESCs after switching 2iL to 2iLA at different time points. b, Western blotting for Erk1/2 in WT , Erk2 -/- , Erk1/2 mutant, Erk2-Res1 or Erk2-Res2 ESCs in 2iL or 2iLA. Erk2-Res: Erk2-rescued in the Erk1/2 mutant ESCs (Lentiviral-based shRNA against Erk1 in the Erk2 -/- ESCs). c-d, qRT-PCR for the Erk1/2 target gene Spry4 (c), and the formative marker genes Wnt8a and Lef1 for WT , Erk2 -/- , Erk1/2 mutant and Erk2-Res ESCs after 4 passages cultured in 2iL or 2iLA(d). e-f, Schematic illustration of the Mek1/2 dKO ESCs generation and the experimental design (e), and Western blotting for validating the deletion Mek1/2 protein of the Mek1/2 dKO ESCs cultured in 2iL or 2iLA (f). g, qRT-PCR for the naive genes Nanog , Prdm14 and formative genes Dnmt3b , Pim2 for WT , Mek1 f/f Mek2 -/- , Mek1/2 dKO ESCs after 5 passages in 2iL or 2iLA. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, t-test. h, Western blotting for the p-Mek1/2 or p-Erk1/2 protein level in Mek1 -/- Mek2 -/- dKO and its control PSCs. i-j, Colony morphology change (i) and qRT-PCR for naive gene Nanog and formative gene Dnmt3l (j) after swtiching 2iL to 2iLA at different time points for WT , M ek1 f/f Mek2 -/- , Mek1/2 dKO, Erk2 -/- or Erk2-Res ESCs.

Article Snippet: The following primary antibodies were used at the indicated dilutions: Zscan4 (Millipore, AB4340); Dnmt3a (Abcam, ab2850), Dnmt3b (Abcam, ab2851), GAPDH (CST, 5174, 1:10,000); Erk1/2 (CST, 4695); p-Erk1/2 (CST, 9101); Nanog (CST, D2A3 XP®).

Techniques: Quantitative RT-PCR, Western Blot, Mutagenesis, shRNA, Marker, Cell Culture, Control

Journal: medRxiv

Article Title: An accurate and rapid Real-time PCR approach for human Monkeypox virus diagnosis

doi: 10.1101/2022.06.23.22276033

Figure Lengend Snippet: Δ Rn vs cycle representation of the qPCR performed on the MIC cycler. MPXV PCR amplification curves from successive 10-fold serial dilutions of a positive sample with Ct values ranging from 15.3 to 22.3.

Article Snippet: PCR amplification was conducted on Mic qPCR (Bio Molecular systems) and AB™7500 Fast Real-Time PCR system (Applied Biosystems, USA) devices.

Techniques: Amplification

Journal: medRxiv

Article Title: An accurate and rapid Real-time PCR approach for human Monkeypox virus diagnosis

doi: 10.1101/2022.06.23.22276033

Figure Lengend Snippet: Δ Rn vs cycle representation of the qPCR performed on the QuantStudio™7 Flex Real-Time PCR System. MPXV PCR amplification curves from successive 5-fold serial dilutions of a positive sample with Ct values ranging from 16.5 to 37.3.

Article Snippet: PCR amplification was conducted on Mic qPCR (Bio Molecular systems) and AB™7500 Fast Real-Time PCR system (Applied Biosystems, USA) devices.

Techniques: Real-time Polymerase Chain Reaction, Amplification