Journal: Cell Death Discovery

Article Title: Combination of eribulin and anlotinib exerts synergistic cytotoxicity in retroperitoneal liposarcoma by inducing endoplasmic reticulum stress

doi: 10.1038/s41420-024-02103-2

Figure Lengend Snippet: A – C Dot plots showing the top 3 KEGG pathway enrichment results of anlotinib, eribulin, and combination group. D GSEA plot featuring protein processing in endoplasmic reticulum pathway. E ERS induced by combination therapy was validated through western blot, wherein key ERS markers were upregulated and apoptosis pathway was initiated. F Rescue experiments further validated that a synergistic effect was largely induced by ERS. After applying salubrinal, synergy scores were significantly reduced.

Article Snippet: Salubrinal was purchased from MedChemExpress (Monmouth Junction, NJ, USA).

Techniques: Western Blot

Journal: Development (Cambridge, England)

Article Title: Mitochondrial dysfunction in oocytes of obese mothers: transmission to offspring and reversal by pharmacological endoplasmic reticulum stress inhibitors.

doi: 10.1242/dev.114850

Figure Lengend Snippet: Fig. 2. Ovulation rate declines with increasing body weight but is improved by treatment with ER stress inhibitors. (A) Hematoxylin and Eosin-stained ovary sections from 14-week-old gonadotropin-treated lean (+/+ or +/bbb) and obese Blobby (bbb/bbb) mice. Arrows indicate unruptured follicles, which are prevalent in ovaries from obese mice. Representative examples from n=4 mice per genotype are shown. Panels a-c are higher magnifications of the indicated follicles of the Blobby mouse. (B) The number of ovulated oocytes in 14-week-old lean and obese mice treated with saline vehicle (Veh.), salubrinal (Sal) or BGP-15 i.p. once per day for 4 days. Values are means+s.e.m. (lean mice+Veh, n=19; lean+Sal, n=5; lean+BGP-15, n=10; obese mice+Veh, n=22; obese+Sal, n=16; obese+BGP-15, n=14). Different letters indicate significant differences calculated by one-way ANOVA, Tukey’s post hoc test; P<0.0001. Right panel: the number of ovulated oocytes in oviducts is not reduced in bbb/bbb mice prior to the onset of obesity, at 6 weeks of age (+/+; +/bbb: n=11; bbb/bbb n=4; Student’s t-test).

Article Snippet: Salubrinal (Invitrogen) was injected i.p. at 1 mg/kg of bodyweight (Sokka et al., 2007) in approximately 0.1 ml saline daily for four consecutive days starting the day prior to treatment with eCG.

Techniques: Staining, Saline

Journal: Development (Cambridge, England)

Article Title: Mitochondrial dysfunction in oocytes of obese mothers: transmission to offspring and reversal by pharmacological endoplasmic reticulum stress inhibitors.

doi: 10.1242/dev.114850

Figure Lengend Snippet: Fig. 5. Treatment with salubrinal or BGP-15 induces mtDNA replication and normalizes mitochondrial membrane potential (ΔΨm) and autophagy in oocytes from obese mice. Obese mice (bbb/bbb) or lean littermates (+/+ or +/bbb) were treated with vehicle (Veh), salubrinal (Sal) or BGP-15 i.p. once daily for 4 days, and oocytes were collected from the oviducts. (A) Live oocytes stained with JC-1, where red fluorescence indicates high ΔΨm, and green indicates low ΔΨm. (B) Ratio of red to green fluorescence, an indicator of mitochondrial activity. Data are presented as means+s.e.m., n=6-30 oocytes from four mice per group. (C) Live oocytes were assessed for autophagic vacuoles, visualized as green fluorescence. Hoechst 33342 (blue). (D) Autophagy levels were quantified as the sum total of green fluorescence within each oocyte. Data are presented as means+s.e.m., n=8-10 oocytes from three mice per group. (E) mtDNA copy number in individual oocytes collected from treated mice. Data are presented as means+s.e.m.; n=19-38 oocytes from six mice per group over three independent experiments. (F) Representative examples of oocytes from three or four mice per treatment group immunostained for TFAM or DRP1 (green) and Hoechst 33342 (blue). Different letters indicate significant differences calculated by one-way ANOVA, Tukey’s post hoc test, P<0.0001.

Article Snippet: Salubrinal (Invitrogen) was injected i.p. at 1 mg/kg of bodyweight (Sokka et al., 2007) in approximately 0.1 ml saline daily for four consecutive days starting the day prior to treatment with eCG.

Techniques: Membrane, Staining, Fluorescence, Activity Assay

Journal: Development (Cambridge, England)

Article Title: Mitochondrial dysfunction in oocytes of obese mothers: transmission to offspring and reversal by pharmacological endoplasmic reticulum stress inhibitors.

doi: 10.1242/dev.114850

Figure Lengend Snippet: Fig. 6. Oocytes from obese Blobby mice exhibit impaired embryo development that is restored by treatment with salubrinal or BGP-15. Obese Blobby (bbb/bbb) mice or lean littermates (+/+ or +/bbb) at 14 weeks of age (except where indicated) were treated with vehicle (Veh; saline), salubrinal (Sal) or BGP-15 daily for 4 days. Ovulated oocytes were collected from oviducts 16 h following hCG injection and fertilized in vitro. (A) At 4 h after in vitro fertilization, many degenerate oocytes (arrows) from untreated obese mice were observed. (B) The percentage of viable oocytes presented as means+s.e.m. Embryo development was assessed on day 2 and day 5 following IVF and is presented as the mean percentage of embryos exhibiting appropriate (‘on-time’) development+s.e.m. (C) Two-cell embryos developed from viable oocytes by day 2. (D) Blastocysts developed from two-cell embryos by day 5. Lean, n=24; lean+Sal, n=7; lean+BGP- 15, n=6; Blobby, n=15; Blobby+Sal, n=6; Blobby+BGP-15, n=7 pools of oocytes from animals within same group. [C,D, right-hand panels are identical assessments of oocytes from 6-week-old (non-obese) mice. +/+, n=13; Blobby, n=10.] (E) Blastocysts were collected at day 5 and the mtDNA copy number of individual blastocysts was measured by using quantitative PCR. Data are expressed as mean copy number+s.e.m.; lean, n=22 blastocysts; lean+Sal, n=10 blastocysts; lean+BGP-15, n=7 blastocysts; Blobby, n=14 blastocysts; Blobby+Sal, n=10 blastocysts; Blobby+BGP-15, n=7 blastocysts collected from three independent IVF experiments. Different letters indicate significant differences by one-way ANOVA, Tukey’s post hoc test; P<0.0001.

Article Snippet: Salubrinal (Invitrogen) was injected i.p. at 1 mg/kg of bodyweight (Sokka et al., 2007) in approximately 0.1 ml saline daily for four consecutive days starting the day prior to treatment with eCG.

Techniques: Saline, Injection, In Vitro, Real-time Polymerase Chain Reaction

Journal: Development (Cambridge, England)

Article Title: Mitochondrial dysfunction in oocytes of obese mothers: transmission to offspring and reversal by pharmacological endoplasmic reticulum stress inhibitors.

doi: 10.1242/dev.114850

Figure Lengend Snippet: Fig. 7. Altered fetal outcomes of oocytes from obese Blobby mice are alleviated after treatment with salubrinal or BGP-15. Obese mice (bbb/bbb) and lean littermates (+/+) were treated with vehicle (Veh; saline), salubrinal (Sal) or BGP-15 for 4 days, followed by the isolation of oocytes from oviducts and fertilization by using IVF. Blastocysts were collected at day 5 of culture and transferred to non-obese pseudo-pregnant recipient mice with six blastocysts per uterine horn. On embryonic day 14.5, fetuses were collected and weighed (A). Liver (B), heart (C) and kidney (D) were dissected and the relative mtDNA copy number normalized to β-actin nuclear DNA was determined. Data are presented as means+s.e.m.; lean+Veh, n=18 fetuses from six surrogates; Blobby+Veh, n=20 fetuses from six surrogates; Blobby+Sal, n=22 fetuses from six surrogates; Blobby+BGP-15, n=15 fetuses from four surrogates. (E) Liver mtDNA samples (n=4 fetuses from each treatment group) were analyzed by using next generation sequencing, and rearrangements were identified. Different letters indicate significant differences calculated by one-way ANOVA, Tukey’s post hoc test; P<0.01.

Article Snippet: Salubrinal (Invitrogen) was injected i.p. at 1 mg/kg of bodyweight (Sokka et al., 2007) in approximately 0.1 ml saline daily for four consecutive days starting the day prior to treatment with eCG.

Techniques: Saline, Isolation, Next-Generation Sequencing