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Fig. 1 | BMC Biology

Fig. 1

From: Combined flow cytometry and high-throughput image analysis for the study of essential genes in Caenorhabditis elegans

Fig. 1

Characterisation of phb-2(tm2998) mutants. a Western blot showing PHB protein levels. In homozygous phb-1(tm2571) and phb-2(tm2998) mutants, both proteins, PHB-1 and PHB-2, are undetectable. Actin is used as loading control. Asterisk denotes unspecific band. One representative western out of three is shown. b Average duration of development for wild-type (n = 12) and phb-2(tm2998) mutants (n = 18) at 20 °C, using the LUC::GFP bioluminescent reporter. Red represents the molts as inferred by low LUC signal. Error bars represent the standard deviation (SD) of the duration of each interval. The graphs below represent the duration of larval stages (L1–L4) and molts (M1–M4) normalised to wild type. The duration of all larval stages (L1–L4) is significantly different between wild type and phb-2(tm2998) mutants (P value < 0.001; two-tailed unpaired t test). The duration of the M2–M4 molting cycles is significantly different amongst wild type and phb-2(tm2998) mutants, with the exception of M1 (P value < 0.001; two-tailed unpaired t test). A representative experiment of two independent replicas is shown. c Lifespans of phb-1(tm2571) (mean = 17±1 days, n = 215) and phb-2(tm2998) (mean = 14.5±0.5 days, n = 302) are significantly shorter than that of the wild type (mean = 18 days, n = 137) (P value < 0.0001, log-rank (Mantel-Cox)). Average of two independent assays is shown. d Background-dependent induction of UPRmt reporters in prohibitin deletion mutants. Fluorescent microscopy images of transgenic animals phb-2(tm2998);Phsp-6::GFP and phb-2(tm2998);Phsp-60::GFP treated with RNAi against the UPRmt components ATFS-1, DVE-1, HAF-1 and UBL-5. Graphical representation of quantification of Phsp-6::GFP (bottom panel, left) and Phsp-60::GFP (bottom panel, right). The induced UPRmt in prohibitin deletion mutants is suppressed upon depletion of atfs-1 and dve-1, whereas the expression of both UPRmt reporters is further increased upon depletion of haf-1 and ubl-5. n > 30 in all conditions. (Mean ± SD; ***P value < 0.001; analysis of variance (ANOVA) test.) One independent replicate out of three is shown

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