Dissecting the membrane cholesterol requirement for mycobacterial entry into host cells

•Optimum host membrane cholesterol is important for the entry of mycobacteria.•Depletion of host membrane cholesterol reduces entry of Mycobacterium smegmatis in macrophages.•Membrane cholesterol replenishment restores entry of M. smegmatis.•Sequestration of membrane cholesterol is sufficient to inhibit M. smegmatis entry.•Enrichment of membrane cholesterol inhibits entry of M. smegmatis into macrophages.