Journal: iScience

Article Title: Peroxisome-dependent and -independent functions of PEX19 in mouse embryonic stem cells

doi: 10.1016/j.isci.2026.117658

Figure Lengend Snippet: Deletion of Pex19 induces mitotic block in mESCs (A) Deletion of Pex19 impairs cell cycle progression and induces polynuclear mESCs. WT, Pex19 KO, and Pex19 KO + PEX19 46C mESCs were stained with PI for DNA content, then subjected to FACS analysis ( n = 3 biological replicates/group; data are presented as mean ± SEM). (B) Deletion of Pex5 in mESCs fails to induce detectable defects in the cell cycle. WT, Pex5 KO, and Pex19 KO 46C mESCs were stained with DAPI for DNA content, then subjected to FACS analysis. (C) Deletion of Pex19 but not Pex5 induces a new fraction of polynuclear mitotic cells. WT, Pex5 KO, and Pex19 KO 46C mESCs treated with or without 50 ng/mL nocodazole were stained with an anti-H3pS10 antibody for detection of mitotic cells and DAPI for DNA content, then subjected to FACS analysis. The M-B population represents polynuclear mitotic cells arrested by deletion of Pex19 or treatment with nocodazole. (D and E) Deletion of Pex19 delays cell cycle progression of mESCs. WT and Pex19 KO 46C mESCs treated with 50 ng/mL nocodazole for 5 h were transferred into normal culture medium without nocodazole, and cell cycle progression was analyzed 15 and 25 min later (three groups of cells (80–250 cells/group) from randomly selected fields were analyzed for each time point for both WT and Pex19 KO mESCs; data are presented as mean ± SEM). Scale bars, 10 μm. See also .

Article Snippet: After fixation with 4% paraformaldehyde, cells were processed for immunofluorescence assay with anti-H3pS10 antibody and phalloidin (Cytoskeleton, PHDH1) staining.

Techniques: Blocking Assay, Staining, Analysis

Journal: iScience

Article Title: Peroxisome-dependent and -independent functions of PEX19 in mouse embryonic stem cells

doi: 10.1016/j.isci.2026.117658

Figure Lengend Snippet: Pex19 KO mESCs have reduced exit from pluripotency during the early stage of neural differentiation (A) Morphology of WT and Pex19 KO 46C mESCs after three days of in vitro neural differentiation. Scale barss, 100 μm. (B) Deletion of Pex19 delays the reduction of pluripotency genes during early neural differentiation. The expression of indicated genes was analyzed before and after three days of in vitro neural differentiation by qPCR ( n = 3 biological replicates/group; data are presented as mean ± SEM; two-way ANOVA, ∗∗∗q < 0.001, ∗∗∗∗q < 0.0001, n.s., not significant). (C) The protein level of Nanog in WT and Pex19 KO cells after three days of in vitro neural differentiation. Scale bars, 20 μm. (D) Adding back PEX19 restores the differentiation-induced reduction of pluripotency protein in Pex19 KO cells. The levels of indicated proteins were analyzed in WT, Pex19 KO (KO), and Pex19 KO + PEX19 (KO + PEX19) 46C mESCs before and after three days of in vitro neural differentiation by immunoblotting. (E) Treatment with DMSO partially increases G1 phase in Pex19 KO cells after three days of in vitro neural differentiation. WT and Pex19 KO 46C mESCs after three days of in vitro neural differentiation with or without 0.1% DMSO were stained with an anti-H3pS10 antibody for detection of mitotic cells and DAPI for DNA content, then subjected to FACS analysis ( n = 3 biological replicates/group; data are presented as mean ± SEM, two-way ANOVA, ∗q < 0.05, ∗∗q < 0.01, n.s., not significant). (F) Adding back PEX19 or treatment with DMSO partially rescues the differentiation defects in Pex19 KO cells. The levels of indicated genes were analyzed in WT, Pex19 KO (KO), or Pex19 KO + PEX19 (KO + P19) 46C mESCs after three days of in vitro neural differentiation with or without 0.1% DMSO by qPCR ( n = 3 biological replicates/group, data are presented as mean ± SEM, two-way ANOVA, ∗q < 0.05, ∗∗q < 0.01, ∗∗∗q < 0.001, and ∗∗∗∗q < 0.0001). (G) Treatment with DMSO partially restores the pluripotency exit defect in Pex19 KO cells. (H) Treatment with a low dose of nocodazole decreases the G1 population in WT cells after three days of in vitro neural differentiation. WT 46C mESCs before (D0) or after three days of in vitro neural differentiation with or without 12.5 ng/mL nocodazole were analyzed for G1 population as in (E) ( n = 3 biological replicates/group; data are presented as mean ± SEM; one-way ANOVA; ∗q < 0.05, ∗∗q < 0.01, n.s., not significant). (I) Treatment with a low dose of nocodazole delays the reduction of pluripotency genes in WT cells after three days of in vitro neural differentiation. The levels of indicated pluripotency genes were analyzed in WT 46C mESCs treated as in (H) ( n = 3 biological replicates/group; data are presented as mean ± SEM, one-way ANOVA, ∗ q < 0.05, ∗∗ q < 0.01, ∗∗∗ q < 0.001, ∗∗∗∗ q < 0.0001, n.s., not significant). See also .

Article Snippet: After fixation with 4% paraformaldehyde, cells were processed for immunofluorescence assay with anti-H3pS10 antibody and phalloidin (Cytoskeleton, PHDH1) staining.

Techniques: Morphology, In Vitro, Expressing, Western Blot, Staining, Analysis

Journal: Cell Communication and Signaling : CCS

Article Title: Avobenzone induces intestinal injury by targeting ATG14 and impairing mitophagy

doi: 10.1186/s12964-026-03211-5

Figure Lengend Snippet: Rapamycin and ISRIB alleviate avobenzone-induced intestinal epithelial injury in mice. A Diagram depicting the experimental design for assessing the effects of Avo on the small intestine in mice treated with Control, Avo (20 mg/kg), Avo (20 mg/kg) + Rapa (5 mg/kg) and Avo (20 mg/kg) + ISRIB (5 mg/kg) for 2 months. B Representative images of the small intestine are displayed, and the length of the small intestine was quantified. C Representative hematoxylin and eosin ( H & E )-stained sections of small intestinal tissues. Crypt density, villus length, and crypt height were quantified. Scale bar, 100 μm. D Representative immunohistochemical images of Ki-67 expression in intestinal crypts and quantification of Ki-67-positive cells per crypt. For ( C ), 30 well-oriented villus–crypt units were analyzed per mouse. For ( D ), 30 crypts per mouse were quantified. The mean value for each mouse was used for statistical analysis ( n = 6 mice per group). Scale bar, 100 μm. E – F Representative immunofluorescence images and quantification of Occludin ( E ) and ZO-1 ( F ) expression in small intestinal tissues. Five randomly selected fields were analyzed for each mouse, and the mean fluorescence intensity obtained from the five fields was used for statistical analysis ( n = 6 mice per group). Scale bars, 50 μm. Data are presented as means ± SD. Statistical significance was determined by one-way ANOVA, as appropriate. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: Sections were blocked with 5% serum for 30 min and incubated with the primary antibody (rabbit anti-Ki67, Abcam, 1:100) overnight at 4 °C, followed by a secondary antibody (HRP-conjugated anti-rabbit, Invitrogen, 1:200) for 2 h. For Hematoxylin-Eosin (H&E) staining, rehydrated sections were stained with Hematoxylin and Eosin (Beyotime, C0105) for 2 min, followed by washing.

Techniques: Control, Staining, Immunohistochemical staining, Expressing, Analysis, Immunofluorescence, Fluorescence

Journal: Nature Communications

Article Title: Low-intensity pulsed ultrasound-mediated nose-to-brain co-delivery of β-blockers and aPD-L1 enhances glioblastoma immunotherapy

doi: 10.1038/s41467-026-76103-4

Figure Lengend Snippet: a was created by Figdraw. a , b Treatment schedule of the GL261 mouse brain tumor model (by figdraw.com). The detailed treatment sequence and time points. c , d Brain images of each group of mice obtained by T2 magnetic resonance imaging (MRI) with different treatments, where tumors are circled in white dashed lines. e-g Immunohistochemical images of tumor sections were examined at 15 days (Ki67 and PCNA, n = 5). h Survival curves of GBM tumor-bearing mice in different treatment groups ( n = 10). Survival analysis was calculated using the log-rank test. i Body weight changes of GBM tumor-bearing mice in different treatment groups ( n = 10). j , k Evaluation of antitumor effects of different treatments. n = 5 samples per group for data in ( c-f ) and ( i-k ). Data are expressed as the mean ± SD. Statistical significances were determined using one-way ANOVA with Fisher’s LSD post‑hoc test. Comparisons were performed between different groups in ( c-f ) and ( i-k ). Not significant (NS) is P ≥ 0.05, and significant P values are shown. n = 10 samples per group for data in ( h ). Survival analysis was compared using the log-rank test. Source data are provided as a file.

Article Snippet: Immunohistochemical staining utilized PCNA (AC058, PCNA Mouse mAb, Abclonal) and Ki67 (A26755PM, Ki67 Rabbit PolymAb®, Abclonal) antibodies, with technical assistance from Nanjing Youmeng Biotechnology Co., Ltd. Tumor sections underwent immunofluorescence staining using a TUNEL assay kit (Beyotime Biotech, Shanghai, China).

Techniques: Sequencing, Magnetic Resonance Imaging, Immunohistochemical staining