Mars analogue environments are some of the most extreme locations on Earth. Their unique combination of multiples extremes (e.g. high salinity, anoxia and low nutrient availability) make them valuable sources for finding new polyextremophilic microbes, and for exploring the limits of life. Mars, especially at its surface, is still considered to be very hostile to life but it probably possesses geological subsurface niches where the occurrence of (polyextremophilic) life is conceivable. Despite their well-recognized relevance, current knowledge on the capability of (facultative) anaerobic microbes to withstand extraterrestrial/Martian conditions, either as single strains or in communities, is still very sparse. Therefore, space experiments simulating the Martian environmental conditions by using space as a tool for astrobiological research are needed to substantiate the hypotheses of habitability of Mars. Addressing this knowledge gap is one of the main goals of the project MEXEM (Mars EXposed Extremophiles Mixture), where selected model organisms will be subjected to space for a period of 3 months. These experiments will take place on the Exobiology facility (currently under development and implementation), located outside the International Space Station. Such space experiments require a series of preliminary tests and ground data collection for the selected microbial strains. Here, we report on the survivability of Salinisphaera shabanensis and Buttiauxella sp. MASE-IM-9 after exposure to Mars-relevant stress factors (such as desiccation and ultraviolet (UV) radiation under anoxia). Both organisms showed survival after anoxic desiccation for up to 3 months but this could be further extended (nearly doubled) by adding artificial Mars regolith (MGS-1S; 0.5% wt/v) and sucrose (0.1 M). Survival after desiccation was also observed when both organisms were mixed before treatment. Mixing also positively influenced survival after exposure to polychromatic Mars-like UV radiation (200–400 nm) up to 12 kJ m−2, both in suspension and in a desiccated form.