Journal: Cell reports

Article Title: Hepatocyte Period 1 dictates oxidative substrate selection independent of the core circadian clock

doi: 10.1016/j.celrep.2024.114865

Figure Lengend Snippet: (A and C) Schematic of in vitro feed/starve experiment in AML12 cells treated with either adenovirus or siRNA targeting Per1, Bmal1, Clock , or Cry1 . (B) Relative expression level of Per1, Fgf21 , and Pdk4 in AML12 cells treated with Ad-GFP or Ad-sh Per1 , fed or starved; n = 5. (D) Relative expression level of Per1, Fgf21 , and Pdk4 in AML12 cells treated with lipofectamine (Control), si Bmal1 , si Clock , or si Cry1 , fed or starved; n = 3. (E) Schematic of ex vivo primary hepatocyte starvation experiment from Per2 WT and Per2 KO female mice. (F) Relative expression level of Per1, Fgf21 , and Pdk4 in primary hepatocytes isolated from Per2 WT or Per2 KO mice, fed or starved; n = 3–4. (G) Schematic of in vivo time-course experiment in Bmal1 fl/fl and Bmal1 cLKO ( Bmal1 fl/fl , Alb-Cre) mice. Mice were either fed ad libitum or fasted for 16 h, and liver tissues were harvested every 4 h in a 24-h duration; n = 3–4. (H) Relative expression level of Per1, Fgf21 , and Pdk4 (normalized to ZT0 Bmal1 fl/fl feed) from liver in (G). (I) Correlation test result between normalized expression level of Per1 and Fgf21 (−8 h or +16 h) (left), Per1 and Pdk4 (−8 h or + 16 h) (middle), and Fgf21 and Pdk4 (right) in (H). Dotted line denotes the 95% confidence interval for the simple linear regression calculation. (J) Schematic of the 14 h + 2 h fast/refeed experimental design in Bmal1 fl/fl ( Bmal1 fl/fl , AAV8-TBG-GFP) and Bmal1 iLKO ( Per1 fl/fl , AAV8-TBG-Cre) mice; n = 3–5. (K) Serum glucose (left), NEFA (middle), and hepatic TG (right) level from mice in (J). (L) Relative expression level of liver Bmal1, Per1, Fgf21, Pdk4 , and Ppara from mice in (J). Data expressed as mean ± SEM. */ a / # p < 0.05, **/ aa / ## p < 0.01, ***/ aaa / ### p < 0.001, ****/ aaaa / #### p < 0.0001 by two-way ANOVA (B, F, K, L), Student’s t test (D), and Pearson correlation test (I). See also and .

Article Snippet: Mouse : Bmal1 fl/fl , Jackson Laboratory , Strain#007668 RRID:IMSR_JAX:007668.

Techniques: In Vitro, Expressing, Control, Ex Vivo, Isolation, In Vivo

Journal: iScience

Article Title: Core circadian transcription factor Bmal1 mediates β cell response and recovery from pro-inflammatory injury

doi: 10.1016/j.isci.2024.111179

Figure Lengend Snippet:

Article Snippet: (Cg)- Arntl tm1Weit /J ( Bmal1 fl/fl ) , Jackson Lab , B6.129S4; RRID:IMSR_JAX:007668.

Techniques: Purification, Recombinant, Plasmid Preparation, In Situ, Enzyme-linked Immunosorbent Assay, Software, Expressing, Microscopy

Journal: The FASEB Journal

Article Title: Regulation of hepatic autophagy by stress-sensing transcription factor CREBH

doi: 10.1096/fj.201802528R

Figure Lengend Snippet: CREBH is regulated by BMAL1 in activating hepatic autophagic genes under the circadian clock. A) Western blot analysis of CREBH, LC3b, ATG7, ATG2b, ULK1, and P62 in the livers of Bmal1-LKO and fl/fl control mice during representative day and night circadian periods. The liver protein lysates were prepared from pooled liver tissues of Bmal1-LKO and fl/fl mice collected at 9 am or 6 pm (n = 3–4 mice/genotype/time point). Levels of BMAL1 and β-actin were determined as controls. The graphs show rhythmic fold changes of protein intensities or ratios (LC3-II/LC3-I) in the livers of Bmal1-LKO and fl/fl mice during the circadian cycle. The protein signals, determined by Western blot densitometry, were normalized to that of β-actin. Fold change of the protein levels was determined by comparing with that at 9 am. B) Western blot analyses of BMAL1, CREBH, LC3b, ATG7, and ATG2b and GAPDH in Bmal1-knockdown and WT control mouse primary hepatocytes expressing the activated CREBH or GFP control under the circadian clock. Mouse primary hepatocytes were infected with Ad expressing Bmal1 short hairpin RNA, CREBH, or GFP before being subjected to horse serum shock for circadian synchronization. Cell lysates were collected throughout a 32-h circadian cycle for Western blot analyses. The graphs show the rhythmic fold changes of activated CREBH, LC3-II/LC3-I ratios, ATG7, and ATG2b protein levels, determined by Western blot densitometry, in Bmal1-knockdown and WT control hepatocytes expressing CREBH or GFP control. Fold change of the normalized protein levels at each circadian time point was compared with that of the starting circadian time.

Article Snippet: Brain and muscle ARNT-like 1 (Bmal1) flox/flox (fl/fl) mice were purchased from The Jackson Laboratory (007668; Bar Harbor, ME, USA) and crossed with Albumin‐Cre transgenic mice (003574) to obtain the liver‐specific Bmal1 KO (Bmal1-LKO) mouse as previously described in Zheng et al. ( 19 ).

Techniques: Western Blot, Control, Knockdown, Expressing, Infection, shRNA