Are you permeabilizing the cells e.g with Triton-X.

Hi,

I do not know what happened, but I can see no attached photo.

Some other suggestions, based on the age and health of the cardiomyocyte source + the lab materials you are using:

https://www.semanticscholar.org/paper/Phospholamban%3A-a-crucial-regulator-of-cardiac-MacLennan-Kranias/c9031efafb1547d1879b3a518720da7df6c636b4

Article Accurate quantitation of phospholamban phosphorylation by immunoblot

Article Alterations of protein expression of phospholamban, ZASP and...

Article Analysis of Cardiomyocyte Development using Immunofluorescen...

Article Biology of the cardiac myocyte in heart disease

Article Protein Quality Control and Degradation in Cardiomyocytes

Article High Resolution Systematic Digital Histological Quantificati...

Two interesting articles:

https://www.jmmc-online.com/article/S0022-2828(16)30357-1/abstract

Article Interplay of cell–cell contacts and RhoA/MRTF‐A signaling re...

Gavin Richardson Hi Gavin,

Appreciated for your help. I had permeabilized the cells with 0.4%Triton X100 for 40mins.

Valeria Tananska

Dear Valeria,

Thank you so much for your suggestion!

The photo disappeared, I'm trying to reupload it.

I think you should revise your working prtocol. Maybe, permeabilization works not optimally.