Agar

After being inoculated into LB medium, P. aeruginosa PAO1 was cultivated at 37 °C and 150 rpm until OD₆₀₀ = 1.0. Then, 1 μL of P. aeruginosa PAO1 bacterial medium was inoculated into the center of swarming solid medium (0.8% nutrient broth (BD), 0.5% glucose, 0.3% agar (BD)) and cultured at 30 °C for 16 h. Then, the colonies of the swarming plate were picked and added to the center of the cooled swarming solid medium (0.8% nutrient broth (BD), 0.5% glucose, 0.5% agar (BD)), which had been supplemented with compounds and cultured at 30 °C for 24 h [40 (link)].

One thousand arsenic treated BEAS-2B (miR-100-inhibitor) and BEAS-2B (miR-NC) cells were re-suspended in 1mL of complete medium (DMEM medium with 10%FBS) containing 0.6% agar and were then plated on top of a bottom layer that contains 1.2% agar (BD) with complete medium. Plates were cultureed under normal conditions for 2 weeks, and then cells colony were counted and photographed under a microscope (Nikon).

Lentil (L. culinaris) plants of Canadian cultivars Eston with complete susceptibility to both races of C. lentis, and CDC Robin with partial resistance to race 1 were grown in 4 inch pots containing commercial potting mixture as described previously9 (link). Isolates CT-30 (race 0) and CT-21 (race 1) of C. lentis were routinely maintained on oat-meal agar (OMA) plates containing 30 g blended quick oats (The Quaker Oats Company, Chicago, USA), 8.8 g agar (Becton, Dickinson and Company, Sparks, USA), and 1 L deionized water supplemented with 0.01% chloramphenicol (AMRESCO, Solon, USA). For infection assays, conidia were collected from 10-day-old OMA plates by flooding the sterile water.

The igf1r deficient R‐cell line was isolated from mouse embryos with a targeted disruption of the igf1r gene by Dr. Renato Baserga's group (Sell et al., 1993). pBABE‐Puro retroviral expression vector (#RTV‐001‐PURO) and Platium‐E packaging cell line (#RV‐101) were bought from Cell Biolabs Inc. (San Diego, CA).
Agar (#214220), BrdU (#550891), 7‐AAD (#559925), mouse anti‐IGF1R (#556000), and FITC labeled mouse anti‐BrdU (#347583) was purchased from Becton, Dickinson and Company (San Jose, CA). Polybrene (sc‐134220), antibodies against GAPDH (sc‐25778), cyclin A (SC‐751), cyclin D (SC‐450), cyclin E (SC‐247), CDK2 (SC‐748), and normal mouse IgG (sc‐2025) were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA). IGF‐1R (#3027), pAkt (#9275), pErk (#9101), SUMO‐1 (#4940), CDK4 (#2906), p27 kip1 (#3698), InsR β (#3020), and phospho‐tyrosine (#9411) antibodies were purchased from Cell Signaling Technology (Danvers, MA). Cyclin B1 (ab181593) and CDK1 (ab18) antibodies were provided by Abcam (Cambridge, MA). qRT‐PCR primers for IGF1R (#Hs00609566_m1) and GAPDH (#Mm99999915_g1), puromycin (#A11138), and protein G Dynabeads (#10004D) were provided by Life Technologies (Carlsbad, CA).

All strains (Table 1) were stored at −80°C. Aggregatibacter actinomycetemcomitans was grown on TSBYE medium containing 3% trypticase soy broth, 0.6% yeast extract with/without 1.5% agar (Becton Dickinson and Company), and incubated statically in a 37°C incubator with 5% humidified carbon dioxide.

Arabidopsis thaliana ecotype Col-0 constitutively expressing intracellular Ca²⁺ indicator aequorin (pMAQ2) is a gift from M. Knight and the principles of how the active aequorin is formed can be found in Knight et al. (1991) (link). Arabidopsis plants were grown in 150 mm × 15 mm round Petri dishes in half-strength Murashige and Skoog salts (MS; Gibco), supplemented with 1.5% (w/v) sucrose (Sigma), and 0.8% (w/v) agar (Becton Dickinson) adjusted to pH 6.0 with KOH in controlled an environmental room at 21 ± 2°C. The fluency rate of white light was ∼110 μmol m^-2 s^-1. The photoperiods were 16 h light/8 h dark cycles. Seeds were sterilized with 2.5% plant preservative mixture (Caisson Laboratories) and stratified at 4°C for 3 days in the dark, and then transferred to the growth room.