Regulation of MAPK and Autophagy Signaling

Experiments were performed as previously described [22 (link)]. Antibodies specific for ERK (#4695), JNK (#9252), p38 (#9212), phospho-ERK (Thr202/Tyr204) (#4370), phospho-JNK (Thr183/Tyr185) (#4668), phospho-p38 (Thr180/Tyr182) (#9211), LC3B (#2775S), mTOR (#2983), phospho-mTOR (Ser2448) (#2971), ULK1 (#8054), phospho-ULK1 (Ser555) (#5869), phospho-ULK1 (Ser757) (#14202), phospho-70S6 kinase (Thr389) (#9234), 70S6 kinase(#2708), phospho-4E-BP1 (Thr37/46) (#2855), 4E-BP1 (#9452), AMPK (#2603) and phospho-AMPK (Thr172) (#2531) were purchased from Cell Signaling Technology (Danvers, MA). Antibodies against DUSP5 (#ab200708) were from Abcam (Cambridge, MA) and antibodies against p62 (#PM045) were from MBL Life science. Specific inhibitors U0126 (#HY-12031A), SB203580 (#HY-10256), SP600125 (#HY-12041) and bafilomycin A1 (#HY-100558) were obtained from MCE. Rapamycin (#V900930-1MG) and rose bengal (#330000) were from Sigma. U0216 (10 μM, 24 h), SB203580 (20 μM, 24 h), SP600125 (15 μM, 24 h), bafilomycin A1 (25 nM, 6 h) and rapamycin (100 nM, 12 h) were added to the cell cultures as described in different experiments. The primers for DUSP5 were TGTCGTCCTCACCTCGCTA and GGGCTCTCTCACTCTCAATCTTC.

Free full text: Click here

Hua C., Zheng Q., Zhu J., Chen S., Song Y., van der Veen S, & Cheng H. (2022). Human Papillomavirus Type 16 Early Protein E7 Activates Autophagy through Inhibition of Dual-Specificity Phosphatase 5. Oxidative Medicine and Cellular Longevity, 2022, 1863098.

Publication 2022

phospho-ERK (Thr202/Tyr204) phospho-JNK (Thr183/Tyr185) phospho-mTOR (Ser2448) phospho-ULK1 (Ser555) phospho-ULK1 70S6 kinase phospho-4E-BP1 (Thr37/46) phospho-AMPK (Thr172) rose bengal

4e bp1 Antibodies Bafilomycin a1 Cell cultures Dusp5 Inhibitors Kinase Mtor Primers Rapamycin Rose bengal Sb203580 Sp600125 U0126 Ulk1

Corresponding Organization : Zhejiang University

Top 5 similar protocols

Variable analysis

independent variables

U0216 (10 μM, 24 h)
SB203580 (20 μM, 24 h)
SP600125 (15 μM, 24 h)
Bafilomycin A1 (25 nM, 6 h)
Rapamycin (100 nM, 12 h)

dependent variables

Activation of ERK, JNK, p38, mTOR, ULK1, 70S6 kinase, 4E-BP1, AMPK
Expression of DUSP5, p62, LC3B

control variables

Antibodies specific for ERK, JNK, p38, phospho-ERK, phospho-JNK, phospho-p38, LC3B, mTOR, phospho-mTOR, ULK1, phospho-ULK1, phospho-70S6 kinase, 70S6 kinase, phospho-4E-BP1, 4E-BP1, AMPK, phospho-AMPK, DUSP5, p62

positive controls

None specified

negative controls

None specified

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!