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. 2015 Jan 12;5:772. doi: 10.3389/fpls.2014.00772

Figure 5.

Figure 5

ProDH1 transcript and protein levels are affected in pi3k hemizygous mutant. Twelve-days-old WT and pi3k hemizygous mutant (PI3K/pi3k) seedlings were stressed with 200 mM NaCl (NaCl) or maintained in 0.5× MS (control, C) during 3 h or 24 h. (A) Proline levels were measured in 24 h-stressed seedlings. Results shown are means ± SD of 6 to 10 biological repetitions in two independent experiments. FW, Fresh Weight. (B) Expression levels of P5CS1 and ProDH1 genes by RT-qPCR on cDNA obtained from 3 h to 24 h- treated WT and pi3k hemizygous mutant seedlings. Results expressed as percentage compared to APT1 as a reference gene are means ± SD (n = 3). Letters indicate statistical differences in P5CS1 (blue letter) or ProDH1 (red letter) gene expression in WT and pi3k hemizygous mutant depending on treatment conditions (Two-Way ANOVA test, P < 0.05). (C) Total proteins obtained from 3 h to 24 h-stressed WT and pi3k hemizygous mutant seedlings were separated by SDS-PAGE. Western blots were incubated with specific antibodies directed against AtProDH proteins. Detection of the immunolabelled proteins was done by autoradiography using an ECL+ kit. Membranes were stained with Ponceau Red as control for protein loading (Rubisco).