Journal: Cell reports
Article Title: Thbs1 regulates skeletal muscle mass in a TGFβ-Smad2/3-ATF4-dependent manner
doi: 10.1016/j.celrep.2024.114149
Figure Lengend Snippet: (A) Fold change in the expression of Thbs1 mRNA in soleus following 30 days of microgravity exposure (space flight) compared to control. Microarray dataset GEO: GSE80223 was analyzed from the Gene Expression Omnibus repository. Error bars denote ±SEM from n = 3 biologically independent animals per group; ** p < 0.01 by two-tailed unpaired Student’s t test. (B) RT-qPCR for Thbs1 , Thbs2 , Thbs3 , Thbs4 , and Thbs5 mRNA in the right tibialis anterior (TA) of 12-week-old wild-type (WT) mice subjected to 3 or 6 days (d) of unilateral hindlimb denervation, compared contralateral sham-operated leg. Error denote ±SEM from n = 3–5 biologically independent animals per group. * p < 0.05 by two-tailed unpaired Student’s t test. (C) Western blot for Thbs1 and Gapdh control in right TA of 12-week-old WT mice subjected to 3 or 10 days (d) of unilateral hindlimb denervation, compared to contralateral, sham-operated TA. n = 2 biologically independent animals per time point. (D) Representative immunohistochemistry for endogenous Thbs1 (green) and BiP (red) to visualize the endoplasmic reticulum (ER) on cryo-embedded TA of WT mice subjected to 10 days (d) of denervation compared to sham-operated controls at 12 weeks of age. Nuclei are shown in blue with DAPI. Scale bars represent 50 μm. (E) RT-qPCR for Thbs1 mRNA from TA of 8-week-old mice fed ad libitum or fasted for 48 h. Data are presented as fold expression over fed WT; error bars denote ±SEM from n = 4 biologically independent animals analyzed per group. * p < 0.05 by two-tailed unpaired Student’s t test. (F and G) Western blot for Thbs1 and Gapdh in TA of 8-week-old mice fed ad libitum or fasted for 48 h (F), and in young (12 weeks of age) and old (24 months of age) WT quadriceps (G). (H) Representative immunohistochemistry for endogenous Thbs1 (green), BiP (red), and the nucleus (DAPI, blue) on cryo-embedded 12-week-old (“young”) and 24-month-old (“old”) WT quadriceps. Scale bars represent 50 μm.
Article Snippet: A literature search yielded a publicly available Affymetrix Mouse Genome 430A 2.0 microarray dataset (NCBI GEO Repository, GSE80223) in which Gambara G. et al. compared the gene expression adaptation in soleus from adult C57Bl/n6 mice that were flown in space aboard the BION-M1 biosatelite for 30 days in orbit ( n = 3), and from sex- and age-matched control mice that were housed in standard vivarium cages ( n = 3).
Techniques: Expressing, Control, Microarray, Two Tailed Test, Quantitative RT-PCR, Western Blot, Immunohistochemistry