I have fractionated ribosomes/polysomes using 10-50% sucrose gradient from HEK-293T cells with and without a treatment. I see the typical peaks of Abs260 for 40S, 60S, 80S, and polysomes under both the conditions. But upon the treatment I see that the 80S peak has increased intensity and lesser number of polysome peaks. Moreover, the polysome peaks were of lesser dense as they appeared in earlier fractions compared to the untreated peaks. But the peaks corresponding to the 40S, 60S and 80S appeared in same fractions for both. All the setup for isolation, and fractionation were similar for both the conditions. The question is, does this say anything about translation status. Does the peaks of polysomes at lesser dense fractions mean that the translation is stalled or anything else? Inputs and suggestions are required.

Thanks in advance