Journal: bioRxiv
Article Title: Mechanosensitive ion channel PIEZO1 enhances endometrial decidualization through BECN1-dependent autophagy
doi: 10.1101/2025.08.05.668790
Figure Lengend Snippet: (A) Expression of LC3A/B in artificial decidualized uteri of Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice 2 or 4 days after stimulation; (B) Representative TEM images of 2-day in vitro decidualized mESCs (E 2 +P 4 treatment, EP) isolated from Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice, vehicle was used as control (CON), red arrows represent autophagosomes; (C) The quantification of autophagosomes in B; (D) The number of autophagosomes and autolysosomes in 2-day in vitro decidualized mESCs (E 2 +P 4 treatment, EP) isolated from Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice, detected by LC3-GFP-RFP vector, representative images shown in figure S4A; (E) Expression of LC3A/B in 2-day in vitro decidualized mESCs (E 2 +P 4 treatment, EP) co-treated with Yoda1 (Y), BAPTA-AM (B), KN93 (K), or U0126 (U); (F) Expression of LC3A/B in 2-day in vitro decidualized HESCs (MPA+cAMP treatment, Mc) co-treated with Yoda1 (Y), BAPTA-AM (B), KN93 (K), or U0126 (U); (G) Expression of p-BECN1 in artificial decidualized uteri of Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice 2 or 4 days after stimulation; (H) Expression of p-BECN1 in 2-day in vitro decidualized mESCs (E 2 +P 4 treatment, EP) co-treated with Yoda1 (Y), BAPTA-AM (B), KN93 (K), or U0126 (U); (I) Expression of p-BECN1 in 2-day in vitro decidualized HESCs (MPA+cAMP treatment, Mc) co-treated with Yoda1 (Y), BAPTA-AM (B), KN93 (K), or U0126 (U); (J) mRNA expression of decidualization marker Prl8a2 in 2-day in vitro decidualized, scrambled (siScra) or Beclin1 siRNA (siBeclin1) transfected mESCs (E 2 +P 4 treatment) isolated from Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice, co-cultured with or without Yoda1; (K) mRNA expression of decidualization marker Prl8a2 in 2-day in vitro decidualized mESCs (E 2 +P 4 treatment) isolated from Piezo1 f/f ( P1 f/f ) and Piezo1 d/d ( P1 d/d ) mice, co-cultured with different dose of Tat-BECLIN1; (L) mRNA expression of decidualization marker genes in 2-day in vitro decidualized HESCs (MPA+cAMP treatment), co-cultured with different doses of Tat-BECLIN1; Scale bar = 500 nm; ns p>0.05; *p<0.05; **p<0.01; ***p<0.001; ****p<0.0001.
Article Snippet: The primary antibodies used in this study included phosphorylated CaMKII (pCaMKII, 1:1000, af3493, Affinit), phosphorylated ERK1/2 (p-ERK1/2, 1:1000, 4370, Cell Signaling Technology), total ERK1/2 (t-ERK1/2, 1:1000, 4695, Cell Signaling Technology), Cyclin D1 (1:1000, 2978, Cell Signaling Technology), CDK4 (1:1000, 23972, Cell Signaling Technology), LC3A/B (1:1000, 4108, Cell Signaling Technology), phosphorylated BECN1 (p-BECN1, 1:1000, PA5-112018, Thermo Fisher Scientific), total BECN1 (t-BECN1, 1:1000, 3495s, Cell Signaling Technology), phosphorylated mTOR (p-mTOR, 1:1000, 2971, Cell Signaling Technology), TUBULIN (1:1000, 2144 S, Cell Signaling Technology), and GAPDH (1:1000, sc-32233, Santa Cruz Biotechnology).
Techniques: Expressing, In Vitro, Isolation, Control, Plasmid Preparation, Marker, Transfection, Cell Culture