Cell Cycle Analysis of hMPCs

The proportion of S phase cells was quantified as previously described (59 (link),75 (link)). Briefly, 5 × 10⁵ digested BMAL1^+/+ and BMAL1^–/– hMPCs were digested by TrypLE™ Express Enzyme (1×) (Thermo Fisher Scientific) and then centrifuged at 1,000 rpm for 5 min. The supernatant was removed and the pellets were fixed in 70% ethanol at −20°C overnight. On the next day, the fixed pellets were washed by PBS and then incubated in PBS containing 0.1% Triton X-100, 0.2 mg/ml RNase A (B100675-0500, Sangon biotech) and 0.02 mg/ml propidium iodide (P3566, Molecular Probes) at room temperature for 30 min. After incubation, the proportion of S phase cells was then assayed by FACS (BD FACS Calibur). Each group has three biological replicates.

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Liang C., Ke Q., Liu Z., Ren J., Zhang W., Hu J., Wang Z., Chen H., Xia K., Lai X., Wang Q., Yang K., Li W., Wu Z., Wang C., Yan H., Jiang X., Ji Z., Ma M., Long X., Wang S., Wang H., Sun H., Belmonte J.C., Qu J., Xiang A.P, & Liu G.H. (2022). BMAL1 moonlighting as a gatekeeper for LINE1 repression and cellular senescence in primates. Nucleic Acids Research, 50(6), 3323-3347.

Publication 2022

TrypLE™ Express Enzyme (1×) RNase A P3566 BD FACS Calibur

Biological Enzyme Ethanol Molecular probes Pellets Propidium iodide Rnase Triton x 100

Corresponding Organization :

Other organizations : Institute of Zoology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Institute of Biophysics, Sun Yat-sen University, Beijing Institute of Genomics, Shanghai Center For Bioinformation Technology, Eighth Affiliated Hospital of Sun Yat-sen University, Capital Medical University, Chinese Academy of Medical Sciences & Peking Union Medical College, Peking Union Medical College Hospital, Renji Hospital, Chinese University of Hong Kong, Salk Institute for Biological Studies

Top 5 similar protocols

Variable analysis

independent variables

BMAL1 genotype (BMAL1+/+ and BMAL1-/-)

dependent variables

Proportion of S phase cells

control variables

Cell type (hMPCs)
Cell handling (digestion by TrypLE™ Express Enzyme, centrifugation, fixation in 70% ethanol, incubation in PBS with Triton X-100, RNase A, and propidium iodide)
FACS analysis

controls

No positive or negative controls were explicitly mentioned in the provided information.

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!